Developmental morphology of the neonatal alligator (Alligator mississippiensis) ovary

American alligator (Alligator mississippiensis) ovary development is incomplete at hatching. During the months following hatching, the cortical processes of oogenesis started in ovo continues and folliculogenesis is initiated. Additionally, the medullary region of the gonad undergoes dramatic restructuring. We describe alligator ovarian histology at hatching, 1 week, 1 month, and 3 months of age in order to characterize the timing of morphological development and compare these findings to chicken ovary development. At hatching, the ovarian cortex presents a germinal epithelium containing oogonia and a few primary oocytes irregularly scattered between somatic epithelial cells. The hatchling medulla shows fragmentation indicative of the formation of lacunae. By 1 week of age, oocytes form growing nests and show increased interactions with somatic cells, indicative of the initiation of folliculogenesis. Medullary lacunae increase in diameter and contain secretory material in their lumen. At 1 month, nest sizes and lacunar diameters continue to enlarge. Pachytene oocytes surrounded by somatic cells are more frequent. Trabeculae composed of dense irregular connective tissue divide cortical nests. Three months after hatching oocytes in meiotic stages of prophase I up to diplotene are present. The ovary displays many enlarged follicles with oocytes in diplotene arrest, thecal layers, lampbrush chromosomes, and complete layers of follicular cells. The medulla is an elaborated complex of vascularized lacunae underlying the cortex and often containing discrete lymphoid aggregates. While the general morphology of the alligator ovary is similar to that of the chicken ovary, the progression of oogenesis and folliculogenesis around hatching is notably slower in alligators. Diplotene oocytes are observed at hatching in chickens, but not until 3 months in alligators. Folliculogenesis is completed at 3 weeks in chickens whereas it is still progressing at 3 months in alligators.     

Postparietal and prehatching ontogeny of the supraoccipital in Alligator mississippiensis (Archosauria, Crocodylia)

The first record of the postparietal bone of Alligator mississippiensis, documented by transverse histological sections, is presented. It is the first evidence of the presence of this bone within Recent reptiles. The postparietal is present in a specimen with a head length of 32.3 mm. The bone is a small dermal plate lying ventrally and posteriorly to the posterior margin of the parietal and dorsally to the trabecular bone, forming a dorsal surface of the supraoccipital portion of the neural endocranium. The trabecular bone develops perichondrally from the dorsal surface of the tectal cartilaginous bridge spanning between the dorsal portions of the otic capsules and occipital pilae. The bridge probably represents the fused tectum synoticum posterior plus tectum posterius. Later in ontogeny, the bridge ossifies endochondrally. The endochondrally ossifying bridge together with its perichondrally ossifying trabecular bone form the future supraoccipital. The trabecular bone is the integral part of the cranial endoskeleton and ontogenetically distinct from the dermal postparietal bone.     

Ontogeny of the partial secondary wall of the otoccipital region of the endocranium in prehatching Alligator mississippiensis (Archosauria, Crocodylia)

The ontogeny of the posterior otic and anterior occipital portions of the neural endocranium of prehatching Alligator mississippiensis was investigated by reconstruction from sectioned material. In Stage 6 of this species, in which the endochondral ossification of the otoccipital region of the neural endocranium is only in its very early stage, two bony outgrowths-laminae-are present at the external wall of the posterior portion of the neural endocranium. The anterior lamina arises from the external surface of the basal plate at the level of the posterior margin of the subcapsular process; the posterior lamina arises from the external surface of that portion of the pila occipitalis that forms the posteroventral wall of the metotic fissure. During ontogeny, both laminae lying in the anteroposterior sequence ossify in membrane, fuse together, grow laterodorsally, and fuse with the lateral wall of the lateral semicircular canal and the crista parotica. This lamina forms a new, secondary wall enclosing the posterior section of the otic capsule and contains the large external jugular foramen (or foramen vagi) in its basal portion. The laminae, designated lamina juxtaotica anterior and posterior (lamina juxtaotica when fused together), have not been recorded previously in crocodylians and are absent in all other Recent reptiles. From the functional point of view, the juxtaotic lamina 1) forms the margins of the external jugular foramen, and 2) forms the floor of the posterior section of the Eustachian tube. In birds, the structure called the metotic cartilage, which arises in ontogeny as an independent element, has a similar position as the juxtaotic lamina. However, the two structures differ in their developmental origins and their relation to the Eustachian tube and the ramus hyomandibularis of the facialis nerve. Moreover, there is no external jugular foramen in birds.     

Ontogeny of the palatoquadrate and adjacent lateral cranial wall of the endocranium in prehatching Alligator mississippiensis (Archosauria: Crocodylia)

The purpose of this article is to gain insight into the ossification sequence of the palatoquadrate and the adjacent lateral cranial wall of prehatching Alligator mississippiensis, a process about which there is almost no published information. Results were obtained by studying serial histological sections of the series of ontogenetic stages and enlarged wax-plate models of several stages. The cartilage of the palatoquadrate starts to ossify endochondrally in the quadrate portion of the pars pterygoquadrata palatoquadrati in Stage 6A. In this stage, a bone, called the lamina palatoquadrati anterior here, appears at and close to the anteromedial wall of the cartilaginous pterygoid portion of the pars pterygoquadrata. The lamina palatoquadrati anterior ossifies in membrane. Later in ontogeny, the lamina palatoquadrati anterior spreads into the cavum epiptericum and sheathes the posterior portion of the trigeminal ganglion laterally. The jaw adductor muscles insert at the outer surface of the lamina palatoquadrati anterior. The lamina palatoquadrati anterior is a new structure not previously recorded in crocodylians or any other Recent reptile. The topology, mode of ossification, and functional anatomy of the lamina palatoquadrati anterior correspond to those of the membranous ossification of the alisphenoid of marsupials. Another bone, called the lamina prootici anterior here, spreads in membrane from the anterolateral wall of the prootic portion of the otic capsule into the prootic fenestra, above the trigeminal ganglion. The lamina prootici anterior represents a structure not recorded previously in crocodylians. It contributes to the orbitotemporal braincase wall.     

扬子鳄与密河鳄基因组DNA的复性动力学和组织结构的研究

通过DNA复性动力学的分析,测定了扬子鳄和密河鳄基因组的组织结构,两者的复性曲线非常相象,高度重复DNA含量很少,而中度重复DNA序列各占基因组的30％左右。中度重复序列部分能再分成三个组分,虽然每个组分的序列复杂性和拷贝数有差别,但每部分的总复杂性是相似的。我们已测得扬子鳄基因组的单拷贝序列为3.68×10’bp,基因组为5.65×10’bp;密河鳄基因组的单拷贝序列为3.67×10’bp,基因组为5.24×10’bp。根据基因组的组织结构的分析,在分子水平上表明两种鳄鱼是亲缘关系十分相近的种。     

Evaluation of Effects of Harvest on Alligator Populations in Louisiana

Biologists with the Louisiana Department of Wildlife and Fisheries, Louisiana, USA, have managed statewide annual harvest of alligators (Alligator mississippiensis) for 35 years (1981–present). We collected and analyzed harvest data for Louisiana alligators to determine the effects of harvest on the population structure, focusing on the larger size classes (≥274 cm) of this slow-growing species. Linear regression analyses revealed that body size-class structure, based on overall average size and the percentage of animals harvested in the larger size classes was relatively stable. Annual aerial alligator nest counts indicated a continual growth of the population, and over time harvested alligators maintained a constant average size. Analyses of population size (based on number of nests and population modeling) indicated that the current annual harvest represents approximately 3% of the population. Linear regression analysis showed that annual hunter success declined only slightly during the study period, and the scheduling of the hunt season after the hatch period and recommended hunting in areas not frequented by breeding females provides economic opportunities for hunters to participate in a sustainable harvest that preserves the larger size classes of alligators in the population. Strict enforcement of existing laws was a key factor responsible for the success of this harvest program. Comparison of alligator population size and number of harvest-related citations indicated that illegal harvest did not have a negative effect on population size, and linear regression analyses revealed that the rate of increase in citations was lower than the increase in populations over the study period. The results of this harvest program indicated that alligators can be hunted in a sustainable manner if hunting is conducted after the hatch period and occurs in areas that primarily exclude the harvest of adult females, and strict law enforcement curbs illegal activities that negatively affect populations. © 2021 The Authors. The Journal of Wildlife Management published by Wiley Periodicals LLC on behalf of The Wildlife Society.     

Identification and comparative analysis of the ovary and testis microRNAome of mud crab Scylla paramamosain

The role of microRNA (miRNA) in reproductive regulation is attracting increasingly more attention. In this study, we obtained 9,643,114 and 15,498,999 raw reads from the ovary and testis library of important farmed mud crab Scylla paramamosain, respectively. After data mining, a total of 4,096,464 and 11,737,973 mappable small RNA sequences remained for analysis. By mapping to the reference genome and expressed sequence tag (EST) of Daphnia pulex and other crabs, a total of 1,417 miRNAs were identified. On the basis of 1,417 miRNAs, 514 (36.3%) unique miRNAs coexpressed in the gonad of female and male libraries, and 336 (23.7%) and 567 (40%) expressed preferentially in female and male libraries, respectively. Analysis of library sequencing data resulted in the identi?cation of 108 miRNAs (out of 1,417; 7.6%) that showed signi?cant differential expression between the two samples. Of these, 13 miRNAs were expressed only in the testis, two miRNAs were expressed only in the ovary, and 93 miRNAs were coexpressed: 57 (61.3%) were upregulated (ovary/testis) and 36 (38.7%) were downregulated (ovary/testis). To confirm the expression patterns of the predicted miRNAs, we randomly selected 14 candidate miRNAs from 108 differentially expressed miRNAs and performed stem–loop real time quantitative PCR (RT‐qPCR) assays in five ovary developing stages. Five miRNAs showed similar expression patterns in almost every stage as those revealed by identification of differentially expressed genes (IDEG6) analysis. The above five miRNAs were predicted to match the 3′‐untranslated region of the published S. paramamosain gene. Four out of five miRNA had a regulation effect on many genes, especially the genes related to gonadal development.     

Studies on skeleton formation in reptiles, v. Patterns of ossification in the skeleton of Alligator mississippiensis DAUDIN (Reptilia, Grocodylia)

Patterns of ossification are described in the endo-and exoskeleton of Alligator mississippiensis. The occurrence of a dermo-supraoccipital is discussed in light of the independence of dermal and endochondral bone. The development of the bony secondary palate is discussed in light of Haeckelian recapitulation. The sequence of ossification in the limb skeleton is shown to differ from the sequence of chondrification of the cartilaginous precursors. Patterns of ossification in Alligator are compared to lepidosaurs in terms of sequence and timing. Important differences relate to ossification patterns in the limb skeleton: lepidosaurs show a dominance of digit III > IV > II > I > V, whereas Alligator shows a dominance of digits III > II> IV > I > V in the ossification process. Ontogenetic repatterning in the ossification of the axial skeleton is discussed as it bears on the serial homology of dorsal ribs, sacral ribs and caudal ribs (transverse processes).     

Cryopreservation of the mouse ovary inhibits the onset of primordial follicle development

The cryopreservation of ovarian tissue has been reported to affect the development of preantral follicles. However, the effect of cryopreservation of ovarian tissue on the development of primordial follicles remains to be elucidated. This study was conducted to evaluate the effect of cryopreservation on the development of frozen-thawed mouse primordial follicles. One-day-old mouse ovaries were cryopreserved by either slow-freezing or a vitrification method. The development of primordial follicles was evaluated histologically and also with markers for follicle development such as: GDF-9, inhibin-alpha subunit and ZP3 in fresh and frozen-thawed ovaries cultured for five days. The proportion of apoptotic and necrotic areas was analyzed in fresh and frozen-thawed ovaries at one and five days after culture, in order to examine the viability of ovarian cells that influence primordial follicle development. The development rate of primordial follicles was significantly lower in slow-frozen and vitrified ovaries than the fresh controls after five days of in vitro culture (P<0.05). The mRNA expression for all developmental markers was slightly decreased in the frozen-thawed ovaries; this difference was not significant. The proportion of apoptosis was significantly increased in the slow-frozen and vitrified ovaries compared to the fresh ovaries at one day (P<0.05); however, there was no difference at five days after culture. The proportion of the area of necrosis was significantly higher in slow-frozen and vitrified ovaries compared to the fresh ovaries at one and five days after culture (P<0.05). Our preliminary data suggest that ovarian tissue cryopreservation using slow-freezing and vitrification methods inhibits development of primordial follicles. This may be caused by the death of ovarian cells through apoptosis and necrosis after cryopreservation.     

扬子鳄胚胎发育分期

本文基于150例不同胎龄的扬子鳄胚胎,将扬子鳄胚胎发育过程分成28个时期。早期胚胎主要以外部形态如体节、体曲程度、脑泡、感觉器官、附肢、鳃弓、颜面部突起、心脏、皮肤等作为分期标准;晚期胚胎主要以器官发育的组织学指标为分期依据。发现在前20个时期中,每个时期的胎龄与密河鳄的很一致,后8个时期的胎龄与密河鳄有差异。分析认为,鳄类晚期胚胎的分期依据应增加器官发育的组织学指标,以便使不同鳄类的胚胎发育有一致的分期标准,为鳄类发育生物学后续研究提供基础资料。     

Differentiation of the gonad rudiment into ovary and testis in the solitary ascidian, Ciona intestinalis

In the early juveniles of Ciona intestinalis, primordial germ cells arise on the degenerated mass of the resorbed tadpole tail, and assemble to form a discrete gonad rudiment. The present study elucidated the morphological sequences during differentiation of the gonad rudiment into the testis and ovary. In 11- to 12-day juveniles, the gonad rudiment, an elongate sac, divided into the testicular and ovarian rudiments. The testicular rudiment separated as a round vesicle from the thickened wall of the elongate sac. The original sac, after separation of the round vesicle, developed into the ovary. In the testicular rudiment, germ cells formed a continuous central mass without association of somatic cells, while in the ovarian rudiment, each germ cell was associated with somatic cells within the epithelium composing the wall of the rudiment. In 13- to 15-day juveniles the testicular rudiment changed into branched tubes ending in club-shaped follicles. Cells characterized by many flattened cisternae of rough endoplasmic reticulum (distal cells) constituted the distal wall of each follicle. Spermatogenic cells were freely present in the follicular lumen, but the largest spermatogonia were in contact with the distal cells. Both in the testicular and ovarian rudiments, germ cells entered meiosis in 18-day juveniles. A novel body (periesophageal body) was found just beneath the ventral margin of the esophageal opening. It comprised irregular follicles made up of one cell type whose cytoplasm, filled with round vesicles and Golgi complexes, was suggestive of an endocrine function. Fragments derived from the periesophageal body were present around the developing ovary.     

超声成像在扬子鳄卵巢等器官形态检测中的应用

扬子鳄(Alligator sinensis)是中国特有的珍稀物种,该物种的生长、繁殖状况一直备受关注,分别在2014年3月(冬眠期)和6月(繁殖期)对17条成年雌性扬子鳄腹部应用VIVIDⅠ型彩色多普勒超声(B型超声)便携诊断仪进行检测,并对3月份卵巢卵泡大小用SPSS 19.0回归分析。扬子鳄心、肝、肠等内脏器官形态在B型超声诊断仪下清晰可见。冬眠期17条扬子鳄两侧卵巢中共检测到41枚卵泡,繁殖期共42枚。冬眠期卵泡处于低回声暗区或无回声暗区的未成熟状态,繁殖期多数卵泡的发育状态与3月相同,少数卵泡呈高回声暗区,此时已形成卵黄颗粒,也有呈高回声亮光区形成卵黄膜的卵泡。对3月份检测的17条扬子鳄具有卵泡的19个卵巢大小与卵泡大小进行回归分析,卵巢随卵泡发育而增大。本研究探讨了B超这种低损伤方法对扬子鳄检测的有效性,同时评估卵泡发育状况,提高繁殖鳄的筛选正确率。     

Possible Generational Effects of Habitat Degradation on Alligator Reproduction

ABSTRACT Population decline of the American alligator (Alligator mississippiensis) was observed in Lake Apopka in central Florida, USA, in the early 1980s. This decline was thought to result from adult mortality and nest failure caused by anthropogenic increases in sediment loads, nutrients, and contaminants. Reproductive impairment also was reported. Extensive restoration of marshes associated with Lake Apopka has been conducted, as well as some limited restoration measures on the lake. Monitoring by the Florida Fish and Wildlife Conservation Commission (FFWCC) has indicated that the adult alligator population began increasing in the early 1990s. We expected that the previously reported high proportion of complete nest failure (θ₀) during the 1980s may have decreased. We collected clutches from alligator nests in Lake Apopka from 1983 to 2003 and from 5 reference areas from 1988 to 1991, and we artificially incubated them. We used a Bayesian framework with Gibbs sampler of Markov chain Monte Carlo simulation to analyze θ₀. Estimated θ₀ was consistently higher in Lake Apopka compared with reference areas, and the difference in θ₀ ranged from 0.19 to 0.56. We conducted change point analysis to identify and test the significance of the change point in θ₀ in Lake Apopka between 1983 and 2003, indicating the point of reproductive recovery. The estimated Bayes factor strongly supported the single change point hypothesis against the no change point hypothesis. The major downward shift in θ₀ probably occurred in the mid-1990s, approximately a generation after the major population decline in the 1980s. Furthermore, estimated θ₀ values after the change point (0.21) were comparable with those of reference areas (0.07–0.31). These results combined with the monitoring by FFWCC seem to suggest that anthropogenic habitat degradation caused reproductive impairment of adult females and decreases in θ₀ occurred with the sexual maturity of a new generation of breeding females. Long-term monitoring is essential to understand population changes due to habitat restoration. Such information can be used as an input in planning and evaluating restoration activities.     

Cytological evaluation of the germ cell development strategy within the testis of the American alligator, Alligator mississippiensis

The cytological changes to germ cells were investigated within the seminiferous epithelium of the American alligator (Alligator mississippiensis). Testicular tissues were collected, embedded in plastic, sectioned on an ultramicrotome, and stained with the periodic acid–Schiff+ procedure followed by a haematoxylin counterstain. Alligators have a prenuptial pattern of germ cell development, where spermatogenesis begins in early spring and sperm is mature by the time mating begins in May. Consistent spatial relationships between germ cells are absent within the seminiferous epithelium of the alligator. Their germ cells progress through the phases of spermatogenesis as a single cohort, leading to one continuous spermiation event that occurs during their mating season (May–June). This temporal germ cell development is different from the consistent spatial development seen within seasonally breeding birds and mammals but is similar to the recently described germ cell development strategies of two other temperate breeding reptiles, the slider turtle and the European wall lizard. The germ cell development strategy shared by these three temperate reptiles representing three different taxa within the class Reptilia is reminiscent of the temporal strategy seen within the anamniotic testis. Thus, alligators and at least two other temperate reptiles exhibit primitive spermatogenic cycles within derived amniotic testes and may be consider intermediates in terms of testicular organization, which may have significance phylogenetically.     

Gonadal sex differentiation in Alligator mississippiensis,a species with temperature-dependent sex determination

Temperature of egg incubation determines sex in Alligator mississippiensis hatchlings. To define the timing and morphology of sexual differentiation, alligator gonads were examined histologically and ultrastructurally throughout embryogenesis. At the male-producing temperature (33° C), the onset of testis differentiation occurred in most embryos during developmental stages 21–22, when a number of somatic cells in the medulla of the gonad became enlarged, forming presumptive Sertoli cells. Some enlarged somatic cells were also observed at the female-producing temperature (30° C) during gonadogenesis, but they were less widespread than at 33° C. Ovarian differentiation at 30° C began slighlty later, during stage 22–23, and was characterised by proliferation of germs cells in the cortex of the gonad. Testis formation in alligators may depend upon presumptive Sertoli cells differentiating prior to a critical event in embryogenesis, such as germ cell proliferation and meiosis. If follows that ovary formation occurs if this requirement is not met, as at lower incubation temperatures.