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Christopher K. Beachy, Kiswendsida C. Ouedraogo Department of Biology, Minot State University, 500 University Avenue West, Minot, North Dakota 58707, USA Cranial development in a miniaturized form of the salamander Desmognathus quadramaculatus:Roles of temperature and thyroid hormone. I-Hypotheses Temperature and Thyroid hormone have an effect on cranial metamorphosis of Desmognathus quadramaculatus II-Background • Description of Plethodontidae salamanders -Variable life cycle -Sensitivity to Temperature and Thyroid hormone • Effect of temperature – Effect on metamorphosis • High temperature effect • Low temperature effect – Structures affected during metamorphosis • External structures: the skin • Internal structures: skeletal elements • Effect of Thyroid hormone – Criteria of TH action • TH concentration • Size-age of the animal III-Materials and Method • Characteristics of the salamanders – Larval size – Place of collection • Gott Farm Creek, Bald Mountains, NC • Experimental Procedure – Number of salamanders – Habitat • Room – – – – Boxes: 8*3*4 LWH 110g sands 100g gravel 250 Ml of distilled water • House : Coolers – Variation in Temperature • Beginning of the experiment – Low temperature (7 C, 11C) • In the course of the experiment – High temperature (11C, 15C) – TH treatment • TH (1.2*10-8 M ) • Control ( 0.0 M) – Variation in the intensity of the treatment factors • Temperature – 1 C every two days • Thyroid hormone – 1.2*10-8 M every two weeks – Four final treatment groups • • • • 11C and 4.8*10-8 M 15 C and 4.8*10-8 M 11 C and control 15 C and control – Evaluation of the internal metamorphosis • The idea of the Bone-Cartilage evaluation – Double-Staining Technique • The idea of the number of teeth and hyobranchial apparatus – Four Dermatocranial elements » Vomer » Parasphenoid » Pterygoid » Maxilla – Hyobranchial Apparatus » Ceratohyal (1) » Basibranchials(2) » Ceratobranchials(2) » Epibranchials (4) Skull of larvae Desmognathus quadramaculatus (dorsal view) Skull of adult Desmognathus quadramaculatus (dorsal view) Skull of adult Desmognathus quadramaculatus (ventral view) • The statistical analysis – Multivariate Analysis of Variance • =0.05 • Test statistics: Wilk’s Lambda – Pearson Correlation • Identification of the nature of the effects of the factors • Identification of the patterns of skeletal elements development IV-Results • Temperature effect – The changes in the hyobranchial apparatus • Changes in the shape • Reduction in the number of structures – The four bones • Vomer – Remodeling of the vomer – Reduction in the number of teeth • Parasphenoid tooth patch – Formation and formation of the teeth • Maxilla – Formation and formation of teeth • Pterygoid – Regression and complete disappearance • Thyroid Hormone – No effect of Thyroid hormone Multivariate Analysis Source Wilk’s F P Temperature 0.11 11.07 <0.001 Thyroxin 0.69 0.64 0.716 Temperature*Thyroxin 0.54 1.22 0.373 Temperature Effect Vomer teeth 400 Teeth patch 350 Pterygoid teeth Number of elements 300 Maxillae teeth 250 200 Totalnumtee 150 Hyobranchial Apparatus 100 masvariatio8 50 score 0 1 Temperature R2 = 0.6564 R2 = 0.5043 R2 = 0.6897 2 2 R2 = 0.2325 R = 0.6545 R = 0.5808 2 Linear (Maxillae teeth) R2=0.601 Linear (Totalnumtee) R2=0.771 Linear (Hyobranchial Apparatus) Linear Summary of Pearson Correlation Correlation 160 Teeth patch Teeth on the other elements 140 120 Pterygoid teeth 100 80 Maxillae teeth 60 40 Totalnumtee 20 0 -20 0 20 40 60 Vomer teeth 2 R = 2 R = 0.7887 2 R = 2 R = 0.0853 V-Discussion • Temperature – Effect of temperature on amphibians – Similarity in the action • Nature of the similarity • Possible conclusion: temperature action is beyond the simple stimulation for regressive or progressive development of skeletal elements • Thyroid Hormone – sensitivity to TH is not related only to metamorphic salamanders. – Insensitivity to TH is not the matter of only non-metamorphic salamanders. – TH may be a secondary factor in metamorphosis • Possible factors: – Mechanical stress: » Internal mechanical stress:Interaction between skeletal elements » External mechanical stress:Action of muscles and feeding system on the skeletal elements – Mechanical stress-inducing factors:Temperature ACKNOWLEDGEMENT Special Thanks to: -The Faculty of the Biology Department of Minot State University and Dr Richard Barkosky -The International Students Office particularly: .Mrs. Ronnie Walker and her family: .Mr. Gary Loper .Beth .Sarah .Davis -Minot Community particularly: .Mr. Tim Eighmy .Mrs. Kathy Hammond .Mrs. Shirley Hintz .Mrs. Margret Lowe .Mr. Berb Williams -Mr. Stuart Melby and Mrs. Karen Melby -Dr David McCormack in the Mathematics Department.