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CELL & TISSUE RESEARCH
Hyperpolarisation-activated and cyclic nucleotide-gated channels are
differentially expressed in juxtaglomerular cells in the olfactory bulb of
mice
Hans-Ulrich Fried, U. Benjamin Kaupp and Frank Müller
Supplementary Information
Identification of immunohistochemical fingerprints (IFs).
IFs were established in two morphologically distinct populations of cells. The first
population included cells without thick dendritic tuft (PG- and SA-like cells), the second
population included cells with a thick dendritic tuft (ET-like cells). In our analysis, we
assumed that all cells in the GL are stained by at least one of our antibodies (see Fig. 6).
Combinations of double and triple stainings were used to establish IFs. We could base our
classification of IFs on combinations of double and triple stainings, because our initial
experiments revealed that markers or HCN isoforms were observed in only few of all
theoretically possible combinations of co-localization and that certain combinations of
markers or HCN isoforms never occurred.
In the following, we provide the experimental strategy to identify each IF. Note that
only unequivocal staining differences (immunonegative versus immunopositive or weakly
immunopositive versus very strong immunopositive) were used to establish the IFs. To keep
the number of figures and panels reasonable, we show only positive staining results in Fig. 4,
suppl. Fig. 4, and suppl. Fig. 5.
1
Experiments A – G define IFs in PG- and SA-like populations
Experiment A: IF1
Initial experiments showed that strong HCN1 was never co-localized with TH, CCK,
PV, CR, HCN2, or HCN3 in PG- or SA-like cells.
HCN1/CB stainings showed that all HCN1-positive cells are either strongly HCN1positive (experiment A) or HCN1- and CB-double positive (experiment D). Thus, only the
following triple stainings were needed to characterize the strong HCN1-positive cells:
HCN1/HCN4/CB stainings (suppl. Fig. 4 a-panels) showed that all strong HCN1positive cells are HCN4- and CB-negative.
HCN1/NOS/vilip1 stainings (suppl. Fig. 4 b-panels) showed that all strong HCN1positive cells are NOS- and vilip1-positive.
Finally, HCN1/NOS/hippo stainings (suppl. Fig. 4 c-panels) showed that all strong
HCN1-positive cells are NOS- and hippo-positive (hippo staining intensity of +/-M, see Table
2).
Conclusion: all strongl HCN1-positive PG- or SA-like cells co-express NOS, vilip1,
and hippo: IF1
Experiment B: IF2
Initial experiments showed that HCN4 was never co-localized with TH, CCK, vilip1,
PV, CR, or HCN2 in PG- or SA-like cells. Thus, only the following triple stainings were
needed to characterize IF2:
HCN1/HCN4/CB stainings
(suppl. Fig.4 a-panels) show:
These cells include IF2 and IF10
HCN4: + to ++ cells
are HCN1: - to +/and CB: - to +/-
2
HCN4: + cells
are HCN1: - or +
and CB: ++ to +++
These cells include IF5 and IF6
HCN4-positive cells can be subdivided into strongly CB-positive and CB-negative (+/-)
populations. All HCN4-pos. / CB-neg. cells are HCN1-negative (- to +/-).
HCN4/CB/hippo stainings
(data not shown) show:
All HCN4-pos./CB-negative cells are also hippo-negative
HCN4/CB/NOS stainings
(data not shown) show:
HCN4: + to ++
CB:
- cells
are NOS: -
HCN4: +
CB: +/- cells
are NOS: +++
These cells are IF10
HCN4-pos. / CB-neg. cells can be subdivided into NOS-negative and strongly NOSpositive populations.
HCN4/NOS/HCN3 stainings
(data not shown) show:
All HCN4-pos./NOS-negative cells are HCN3-negative
Conclusion: among the HCN4-positive PG-SA-like cells, there is one subpopulation
that expresses only HCN4, but no other markers or HCN isoforms tested: IF2
Experiment C: IF3 and IF4
Initial experiments showed that CCK was never co-localized with TH, NOS, hippo,
CR, CB, HCN1, HCN2, or HCN4 in PG- or SA-like cells. Thus, only the following triple
stainings were needed to characterize IF3 and IF4:
Stainings with antibodies directed against HCN1/CCK/ and either vilip1 (suppl. Fig. 4
d-panels) or HCN3 (data not shown) showed that all CCK-positive cells are HCN1-negative
3
but vilip1- and HCN3-positive. The presence of HCN3 in IF3 and IF4 is shown in suppl. Fig.
4 e-panels and g-panels, respectively.
Finally, HCN1/CCK/PV stainings (suppl. Fig. 4 f-panels) showed that CCK-positive
cells are either PV-negative (IF3) or PV-positive (IF4).
Conclusion: CCK-positive PG- or SA-like cells co-express vilip1 and HCN3, but
segregate into PV-positive (IF4) and PV-negative (IF3) populations.
Experiment D: IF5 and IF6
Initial stainings showed that CB was never co-localized with TH, CCK, NOS, vilip1,
PV, CR, HCN2, or HCN3 in PG- or SA-like cells. Thus, only the following stainings were
needed to characterize IF5 and IF6:
CB/hippo stainings (suppl. Fig. 4 i-panels) showed that all CB-positive cells are also
hippo-positive.
CB/HCN1/HCN4 stainings (suppl. Fig. 4 a-panels) showed that all CB-positive cells
are HCN4-positive but can be subdivided in HCN1-positive and HCN1-negative populations.
Conclusion: CB-positive PG- or SA-like cells co-express hippo and HCN4, but
segregate into HCN1-positive (IF5) and HCN1-negative (IF6) populations.
Experiment E: IF7 and IF8
Initial experiments showed that CR was never co-localized with TH, CCK, hippo, PV,
CB, HCN1, HCN2, HCN3, or HCN4 in PG- or SA-like cells. Thus, only the following triple
staining was needed to characterize IF7 and IF8:
CR/NOS/vilip1 stainings (suppl. Fig. 4 h-panels) showed that CR-positive PG- or SAlike cells can be subdivided in two population: IF7 is negative for NOS and vilip1, IF8 is
positive for NOS and variable for vilip1.
4
Experiment F: IF9
Initial experiments showed that TH was never co-localized with CCK, NOS, vilip1
PV, CR, CB, HCN1, HCN2, HCN3, or HCN4 in PG- or SA-like cells. Thus, only a TH/hippo
double staining was needed to characterize IF9:
TH-positive PG- or SA-like cells (IF9) express hippo to a variable degree (- to ++).
Note that, as varibility in hippo staining was very large, we did not subclassify cells into
hippo-positive and hippo-negative cells.
Experiment G: IF10
Inital experiments showed that NOS was never co-localized with TH, CCK, PV, CB,
or HCN2 in cells with PG- or SA-like morphology. Thus, only the following triple stainings
were needed to characterize IF10:
HCN1/HCN4/NOS stainings
(Fig.4b to e) show:
NOS: +++ cells
are HCN1: +++
and HCN4: -
NOS: + to +++ cells
are HCN1: and HCN4: -
These cells are
IF1
These cells are
IF8
NOS: +++ cells
are HCN1: +/and HCN4: +
Three different populations of cells express NOS. From experiments “A“ and “E“, we
know that two of the NOS-pos. populations (IF1 and IF8) do not express HCN3.
5
NOS/HCN3 stainings
(Fig. 4c to h) show:
These cells include IF1 and IF8
NOS: + to +++
HCN3: -
NOS: +++
HCN3: ++
HCN3/CR and
HCN3/hippo stainings
(data not shown) show:
All HCN3-pos. cells are CR- and hippo-negative
CR/NOS/vilip1 stainings
(suppl. Fig.4 h-panels) show:
These cells are IF1
NOS: +++
CR: - cells
are vilip1: +
NOS: +++
CR:
- cells
are vilip1: -
NOS-pos. / CR-neg. cells can be subdivided in vilip1-positive and vilip1-negative
populations. From experiment “A“, we know that the vilip1-positive population must be IF1.
Conclusion: NOS-positive PG- or SA-like cells fall into three populations (IF1, IF8,
and IF10). NOS, HCN3, and HCN4 are exclusively co-localized in IF10. Both IF1 and IF8 do
not express HCN3 or HCN4.
Experiment G: IF11
HCN2 was not co-localized with any of our markers or other HCN isoforms in PG- or
SA-like cells. Thus, HCN2 labeled a distinct cell population (IF11) that is not detected by any
other marker.
Conclusions of experiments A to G
Using antibodies against HCN1, HCN2, HCN4, CCK, CB, CR, TH, and NOS as
starting points of our analysis, we have identified 11 different IFs in experiments A to G.
Further stainings with antibodies not used as a starting point (i.e. vilip1, hippo, PV, and
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HCN3) revealed no further IFs in PG-and SA-like cells (data not shown). Because, we
showed that our antibodies most likely stain all juxtaglomerular cells, IF1 to 11 are the only
exciting immunohistochemical fingerprints in PG-and SA-like cell. However, we can not
exclude that the IFs defined here can be further subdivided using additonal antibodies.
Experiments I - V define IFs in ET-like populations
Inital experiments showed that cells with ET-like morphology
1. do not express HCN2;
2. are either positive for CCK or for TH (there are no ET-like cells that are either negative or
positive for both CCK and TH).
Experiment I: IF12
HCN1/CCK/NOS stainings
(suppl. Fig.5 a-panels) show:
IF12
CCK: + to ++ cells
are HCN1:+ to ++
and NOS: + to ++
IF13
CCK: + to ++ cells
HCN1: +/NOS: + to ++
IF14
IF15
IF16
CCK: + to ++ cells CCK: + to ++ cells CCK: HCN1: + to ++
HCN1: HCN1: NOS: +/NOS: NOS: +
This triple staining showed five different staining patterns (IF12 to IF16). Only IF12 is
HCN1- and NOS-positive.
Stainings with antibodies against HCN1/NOS/ and either HCN4, hippo, vilip1, or
HCN3 showed that all HCN1- and NOS-positive cells are HCN3- and HCN4-positive but
hippo- and vilip1-negative. Localization of HCN3 and HCN4 in IF12 cells is shown in suppl.
Fig. 5 c-panels and e-panels, respectively.
Conclusion: CCK-positive ET-like cells fall in four populations (IF12, IF13, IF14, and
IF15). IF12 is characterized by co-expression of CCK, NOS, HCN1, HCN3, and HCN4.
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Experiment II: IF13
HCN1/CCK/NOS stainings
(suppl. Fig.5 a-panels) show:
IF12
CCK: + to ++ cells
are HCN1:+ to ++
and NOS: + to ++
IF13
CCK: + to ++ cells
HCN1: +/NOS: + to ++
IF14
IF15
IF16
CCK: + to ++ cells CCK: + to ++ cells CCK: HCN1: + to ++
HCN1: HCN1: NOS: +/NOS: NOS: +
This triple staining showed five different staining patterns (IF12 to IF16). Two
populations (IF13 and IF16) are NOS-positive but HCN1-negative.
HCN1/NOS/vilip1 stainings
(data not shown) show:
All HCN1-neg. / NOS-pos. cells are vilip-negative
HCN1/NOS/HCN3 stainings
(suppl. Fig. 5 b-panels) show:
HCN1: +/NOS: + to ++ cells
are HCN3: +
HCN1: NOS: + cells
are HCN3: +/-
These cells are IF16
HCN1-neg. / NOS-pos. cells can be subdivided into HCN3-positive and HCN3negative populations. For the following reason, the HCN3-positive cells are IF13, while the
HCN3-negative cells are IF16. TH/HCN3 double stainings show that all TH-negative cells
express HCN3. Note that all ET-like cells do either express CCK or TH, TH-negative cells
must be CCK-positive. Therefore, IF13 must be HCN3-positive.
HCN1/NOS/HCN4 stainings
(suppl. Fig. 5 e-panels) show:
HCN1: +/NOS: + to ++ cells
are HCN4: + to ++
HCN1: NOS: + cells
are HCN4: +/-
These cells are IF16
HCN1-neg. / NOS-pos. cells can be subdivided into HCN4-positive and HCN4negative populations. TH/HCN4 double stainings show that only TH-negative cells (thereby
CCK-positive) express HCN4. Thus, IF13 must be HCN4-positive.
8
NOS/HCN4/hippo stainings
(data not shown) show:
All NOS- and HCN4-positive cells are hippo-negative
Conclusion: of the four CCK positive ET-like cell populations, IF13 is characterized
by co-expression of NOS, HCN3, and HCN4.
Experiment III: IF14
HCN1/CCK/NOS stainings
(suppl. Fig.5 a-panels) show:
IF12
CCK: + to ++ cells
are HCN1:+ to ++
and NOS: + to ++
IF13
IF14
CCK: + to ++ cells CCK: + to ++ cells
HCN1: +/HCN1: + to ++
NOS: + to ++
NOS: +/-
IF15
IF16
CCK: + to ++ cells CCK: HCN1: HCN1: NOS: NOS: +
This triple staining showed five different staining patterns (IF12 to IF16). Only IF14 is
HCN1-positive and NOS-negative (around detection threshold).
Stainings with antibodies directed against HCN1/NOS/ and either HCN3, HCN4,
vilip1, or hippo showed that all HCN1-positive and NOS-negative cells are HCN3-, vilip1-,
and hippo-positive but HCN4-negative (suppl. Fig. 5 e-panels). That IF14 cells are HCN3-,
vilip1-, and hippo-positive is shown in suppl. Fig. 5 b-, d-, and g-panels, respectively.
Conclusion: in the four CCK positive ET-like cell populations co-expression of vilip1,
hippo, HCN1, and HCN3 is only seen in IF14.
Experiment IV: IF15
HCN1/CCK/NOS stainings
(suppl. Fig.5 a-panels) show:
IF12
CCK: + to ++ cells
are HCN1:+ to ++
and NOS: + to ++
IF13
IF14
IF15
CCK: + to ++ cells CCK: + to ++ cells CCK: + to ++ cells
HCN1: +/HCN1: + to ++
HCN1: NOS: + to ++
NOS: +/NOS: -
IF16
CCK: HCN1: NOS: +
This triple staining showed five different staining patterns (IF12 to IF16). Only IF13
and IF15 are HCN1-negative and CCK-positive.
9
Stainings with antibodies directed against HCN1/CCK/ and either vilip1, HCN3 or
HCN4 showed that all HCN1-negative and CCK-positive cells are vilip1-negative but HCN3and HCN4-positive. That HCN3 and HCN4 are expressed in IF15 cells is shown in suppl. Fig.
5 f-panels.
Finally, the HCN1/CCK/hippo staining showed that HCN1-negative and CCKpositive cells segregate into hippo-negative and hippo-positive populations. We know from
experiment II that IF13 is hippo-negative, thus, IF15 must be hippo-positive. Conclusion: Of
the four CCK positive ET-like cell populations co-expression of hippo, HCN3, and HCN4 is
only observed in IF15.
Experiment V: IF16 and IF17
TH/hippo/NOS stainings (suppl. Fig. 5 i-panels and j-panels) showed that TH-positive
ET-like cells segregate in NOS-positive (IF16) and NOS-negative (IF17) populations. Both
populations had a variable expression of hippo (staining intensity of – to ++). Further
experiments showed that TH was never co-localized with CCK, vilip1, HCN1.
Suppl. Fig. 1 Specificity of HCN stainings in the GL revealed by double labelings with
antibodies recognizing different epitopes of HCN1 (a to c), HCN2 (d to f), HCN3 (g to h), and
HCN4 (j to l), respectively. Antibodies (see table 1) are specified in each panel. In each of the
double labelings antibodies directed against the same HCN isoform yielded qualitatively
identical staining patterns. This indicates specificity of antibodies with cross-reactivity below
background levels. Scale bars = 10 m
Suppl. Fig. 2 Specificity of HCN1 and HCN3 antibodies revealed in knockout animals. a and
b: Representative stainings with antibodies directed against HCN1 (HCN1) in wild type (a)
10
and HCN1 knockout (b) animals. c and d: Representative stainings with antibodies directed
against HCN3 (HCN3 Shigemoto) in wild type (c) and HCN3 knockout (d) animals. No
staining is observed in the knockout animals indicating specificity of antibodies with crossreactivity below background levels. Scale bars = 100 m
Suppl. Fig. 3 Staining with antibodies directed against hippocalcin (hippo) at lower
magnification. Hippo staining is found in several layers of the olfactory bulb (GL, EPL, IPL,
and GrL). No staining is visible in the AL and ML. EPL, external plexiform layer; GL,
glomerular layer; GrL, granule cell layer; IPL, internal plexiform layer; ML, mitral cell layer;
AL, olfactory nerve layer. Scale bars = 100 m
Suppl. Fig. 4 Immunohistochemical identification of PG-like cell populations. a-panels (a,
a’, a’’, and a’’’): Example of stained PG-like cells in a lower magnification. Triple labeling
for HCN1 (green, HCN1), HCN4 (red, PG2-1A4), and CB (blue, CabPneu) reveals IF1
(green arrowhead, only HCN1 positive), IF2 (blue arrowhead, only HCN4 positive), IF5 (red
arrowhead, HCN1, HCN4, and CB positive), and IF6 (light turquoise arrowhead, HCN4 and
CB positive). b- and c-panels: Immunohistochemical identification of cells with IF1. IF1
cells (green arrowhead) are characterized by the co-localization of HCN1, NOS, vilip1, and
hippo as seen in two triple labelings (b-panels: HCN1 in green, NOS in red, and vilip1 in
blue; antibodies used were: HCN1, NOSmono, and vilip1 c-panels: HCN1 in green, NOS in
red, and hippo in blue; antibodies used were: HCN1 NOSmono, and hippo). d-, e-, f-, and
g-panels: Immunohistochemical identification of cells with IF3 and IF4. Both, IF3 (light
orange arrowhead) and IF4 (dark orange arrow head) feature co-expression of HCN1, HCN3,
CCK, and vilip1. The HCN1 staining is often just above background and therefore not clearly
visible in all stainings. PV is only detectable in IF4. The co-localization patterns in IF3 and
IF4 are shown in four triple stainings (d-panels: HCN1 in green, CCK in red, and vilip1 in
11
blue; antibodies used were: HCN1, CCK, and vilip1; e-panels: CCK in green, HCN3 in red,
and PV in blue; antibodies used were: CCK, HCN3 Shigemoto, and PV f-panels: HCN1 in
green, CCK in red, and PV in blue; antibodies used were: HCN1, CCK, and PV g-panels:
HCN1 in green, HCN3 in red, and PV in blue; antibodies used were: HCN1, HCN3
Shigemoto, and PV). h-panels: Immunohistochemical identification of cells with IF7 and IF8.
Triple labeling for CR (green, CAL), NOS (red, NOSmono), and vilip1 (blue, vilip1) reveals
IF7 (black arrowhead, only CR positive) and IF8 (grey arrowhead, CR, NOS, and vilip1
positive). i-panels: Immunohistochemical identification of cells with IF5 and IF6. Triple
labeling for HCN1 (green, HCN1), hippo (red, hippo), and CB (blue, CabPneu) reveals IF5
(red arrowhead, HCN1, hippo, and CB positive) and PG6 (light turquoise arrowhead, hippo
and CB positive). Note: cells with IF5 and IF6 have also been characterized in the a-panels. jpanels: Immunohistochemical identification of cells with IF9. Co-localization of TH (green,
TH) and hippo (red, hippo) reveals IF9 (white arrowhead, TH and hippo positive). Colour
codes of arrowheads same as in table 2. The a-panels and h-panels are also shown in Fig. 3.
Scale bars = 10 m
Suppl. Fig. 5. Immunohistochemical identification of ET-like cell populations. a-, b-, c-, d-,
e-, f-, g-, and h-panels: Immunohistochemical identification of cells with IF12, IF13, IF14,
and IF15 cells. a-panels: The triple staining for HCN1 (green, HCN1), CCK (red, CCK),
and NOS (blue, NOSmono) reveals four different CCK-positive IFs in ET-like cells. IF12
(white arrows): strong co-expression of HCN1, CCK, and NOS. IF13 (dark purple arrow): coexpression of CCK and NOS, little or no HCN1. IF14 (orange arrow): co-expression of
HCN1, CCK, and only weakly NOS. IF15 (light purple arrow): only CCK expression. bpanels: The triple staining for HCN1 (green, HCN1), HCN3 (red, HCN3 Shigemoto), and
NOS (blue, NOSpoly) shows the expression of HCN3 in cells with IF14 (orange arrow,
12
identified by moderate HCN1 and weak NOS staining). A weak HCN3 expression is also seen
in cells with IF13 (dark purple arrow, identified by strong NOS and no HCN1). c-panels: The
triple staining for HCN1 (green, HCN1), HCN3 (red, HCN3 Shigemoto), and vilip1 (blue,
vilip1) shows the strong expression of vilip1 in cells with IF14 (HCN1 (green) and HCN3
(red) positive) and weak expression of vilip1 in cells with IF12 (also HCN1 (green) and
HCN3 (red) positive). In a vilip1 and NOS double staining the strong vilip1 expression in
weakly NOS-positive cells (IF14) was confirmed (data not shown). d-panels: In the staining
for HCN1 (green, HCN1), CCK (red, CCK), and vilip1 (blue, vilip1), the sensitivity of the
detector for vilip1 was adjusted to match the strong staining in IF14 (orange arrow; note the
pronounced dendritic tuft in the ET like cells shown here). Expression of vilip1 in cells with
IF12 is much weaker and, therefore, not visible at this detector setting. e-panels: The triple
staining for HCN1 (green, HCN1), HCN4 (red, PG2-1A4), and NOS (blue, NOSpoly)
shows that HCN4 is found in IF12 (white arrow, HCN1 and strong NOS expression) and IF13
(purple arrow, strong NOS and no HCN1 expression), but not in IF14 (orange arrow, weak
NOS and strong HCN1 expression). f-panels: A staining with four primary antibodies HCN4
(green, PG2-1A4), HCN3 and NOS (both red, HCN3 Shigemoto and NOSpoly, respectively),
and TH (blue, TH) is shown. Although using the same secondary antibody, due to the
difference in subcellular localization, NOS (cytoplasmic staining) and HCN3 (plasma
membrane staining) could be clearly differentiated. HCN3 and HCN4 are co-expressed in
cells with IF15 (light purple arrow). g-panels: Triple staining for HCN1 (green, HCN1),
HCN4 (red, PG2-1A4), and hippo (blue, hippo). Hippo is present in IF14 (orange arrow,
strong HCN1 and no HCN4 expression). h-panels: Triple staining for HCN1 (green,
HCN1), CCK (red, CCK), and hippo (blue, hippo). Expression of hippo is found in a cell
positive for CCK but negative for HCN1, a feature characteristic for either IF13 or IF15 (light
purple arrow). Because hippo is not found in strongly NOS-positive ET cells (i.e. IF13), the
expression pattern of the cell (CCK and hippo, but no HCN1) is consistent with IF15 (data not
13
shown). The second CCK-positive but HCN1-negative cell in this panel (purple arrow, IF13)
is negative for hippo. i- and j-panels: Immunohistochemical identification of cells with IF16
and IF17. IF16 and IF17 are positive for TH and hippo (IF16, blue arrow; IF17 grey arrow).
The triple staining for TH (green, TH), hippo (red, hippo), and NOS (blue, NOSgt) shows that
all three markers are found in IF16 (blue arrow, i-panels). In a triple staining with the same set
of antibodies, IF17 is positive for TH and hippo but negative for NOS (j-panels). Colour
codes of arrows same as in table 2. The d-, i-, and j-panels are also shown in Fig. 5. Scale bars
= 10 m
Suppl. Fig. 6. Double stainings with antibodies directed against GFAP (green) and HCN
channels (red) show that no HCN staining is colocalized with GFAP. Thus, the majority of
HCN1 (a), HCN2 (b), HCN3 (c), and HCN4 (d) channels are found in neurons rather then
astrocytes. Antibodies used were: GFAP and HCN1(a), GFAP and QQA-1A6 (b), GFAP
and HCN3 (c), and GFAPab and PG2-1A4 (d). Arrows in c point to HCN3 positive cells.
Scale bar = 100 m
Suppl. Fig. 7. 3D reconstructions of confocal z-stacks. a: Reconstructions of two CB positive
(CabPneu) PG-like cells (arrowhead) with one or two dendrites, which ramify within the same
glomerulus. b: Reconstruction of a NOS positive (NOSpoly) SA-like cell (arrow) with a long
reaching dendrite. Three other cells (asterisk) are in direct vicinity of the SA-like cell body or
dendrite and were not perfectly reconstructed as separate objects. c: Reconstruction of three
vilip1 positive PG-like cells (arrowheads) with several dendrites, and an ET-like cell (arrow)
with a thick primary tuft. d: Reconstruction of a NOS positive (NOSpoly) ET-like cell
(arrow) with a tick primary tuft (single asterisk) and a thick lateral dendrite (double asterisk).
Scale bars = 10 m
14
Suppl. Table 1. Number of cells and glomeruli counted to average number of cells per
glomerulus and section in table 3.
IF
IF1
IF2
IF3
IF4
IF5
IF6
IF7
IF8
IF9
IF10
IF11
IF12
IF13
IF14
IF15
IF16
IF17
cells counted across number of glomeruli
717
371
562
410
191
258
522
521
318
180
145
142
95
272
88
36
125
350
190
301
355
190
190
121
121
140
476
343
194
194
301
150
140
140
15