Genital anatomy and copulatory interactions in the broad snouted Caiman (Caiman latirostris).

The broad snouted caiman is a crocodylian native to South America that is subject to extensive conservation management in both wild and farming environments. Although reproductive behaviors like egg laying and clutch care have been examined in this species, little else is known about their copulatory system. We examined the anatomy of male and female cloacal and genital tissues ex vivo to build hypotheses of their interactions during copulation and the effects of that interaction on insemination.

Glans inflation morphology and female cloaca copulatory interactions of the male American alligator phallus†.

The phallic glans of the American alligator (Alligator mississippiensis) is the distal termination of the semen-conducting sulcus spermaticus and during copulation has the closest, most intimate mechanical interactions with the female urodeum, the middle cloacal chamber that contains the opening to the vaginal passages and oviducts. However, the details of this interface leading to insemination and gamete uptake are unclear. Here, we: (1) histologically characterize the underlying tissue types and morphologically quantify the shape changes associated with glans inflation into the copulatory conformation, (2) digitally reconstruct from MRI the 3D shape of functional tissue compartments, and (3) diffusible iodine-based contrast-enhanced computed tomography image the copulatory fit between male phallus and female cloaca.

Low oxygen: A (tough) way of life for Okavango fishes.

Botswana's Okavango Delta is a World Heritage Site and biodiverse wilderness. In 2016-2018, following arrival of the annual flood of rainwater from Angola's highlands, and using continuous oxygen logging, we documented profound aquatic hypoxia that persisted for 3.5 to 5 months in the river channel.

Morphological changes associated with Nile crocodile (Crocodylus niloticus) phallic glans inflation.

The crocodylian phallic glans is the distal inflatable structure that makes the most direct contact with the female cloacal and associated reproductive tract openings during copulation. Therefore, its form and function directly impact female tissue sensory interactions and insemination mechanics. Compared to mammals, less is known about glans functional anatomy among other amniotes, including crocodylians.

Crocodylian conservation and evolution insights from an anatomical and histological examination of phalli from male false gharial (Tomistoma schlegelii).

As wild population threats for the endangered false gharial (Tomistoma schlegelii) persist, conservation breeding programs, including developing semen collection techniques for subsequent artificial insemination, are becoming important species conservation measures. Developing reproductive biology understanding of a species is important to developing best practices and hopefully maximizing reproductive successes. However, information on crocodylians functional copulatory anatomy in general is lacking.

Correlation between increased postpubertal phallic growth and the initiation of cranial sexual dimorphisms in male Morelet's crocodile.

While puberty is an animal commonality, little is known of its timing or process in crocodylians. Males copulate with an intromittent phallus that has a distinct glans morphology which directly interacts with the female cloaca, putatively effecting effective semen transfer and ultimately increased fecundity. Here we present, during the Morelet's crocodile lifecycle, a well-defined body length (65 cm snout-vent length) inflection point that marks a subsequent increase of phallic glans growth rates.

AHR and CYP1A expression link historical contamination events to modern day developmental effects in the American alligator.

The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor that initiates a transcriptional pathway responsible for the expression of CYP1A subfamily members, key to the metabolism of xenobiotic compounds. Toxic planar halogenated aromatic hydrocarbons, including dioxin and PCBs, are capable of activating the AHR, and while dioxin and PCB inputs into the environment have been dramatically curbed following strict regulatory efforts in the United States, they persist in the environment and exposures remain relevant today. Little is known regarding the effects that long-term chronic exposures to dioxin or dioxin-like compounds might have on the development and subsequent health of offspring from exposed individuals, nor is much known regarding AHR expression in reptilians.

The Morphological Diversity of Intromittent Organs: An Introduction to the Symposium.

Intromittent organs are structures that enter the female genital tract and deposit sperm; these organs are found in many animal taxa that use internal fertilization. Despite their shared function, they are fantastically diverse morphologically. Many of their species-specific shape differences are likely the result of sexual selection and coevolution between male and female reproductive tracts, but a growing number of studies have identified other factors that can also affect their functional anatomy.

Morphological characteristics regulating phallic glans engorgement in the American alligator.

The distal part of the crocodilian phallus consists of a bulbous glans containing well-developed vascular tissues that can inflate before or during sexual activity, enlarging and elaborating the glans into a complex, though still functionally undefined, copulatory structure. An enlarged glans putatively interacts with the female cloaca and may change the shape of her reproductive tract to facilitate insemination and increase the probability of fertilization. Here, we investigated the cellular-level properties of the glans and other inflatable phallic tissues associated with the sperm-conducting sulcus spermaticus in the American alligator (Alligator mississippiensis).

Gonadal differentiation in reptiles exhibiting environmental sex determination.

As temperature-dependent sex determination (TSD) and homozygote or heterozygote genetic sex determination (GSD) exist in multiple reptilian taxa, they represent sex determination systems that have emerged de novo. Current investigations have revealed that the genetic mechanisms used by various reptilian species are similar to those used by other vertebrates. However, the recent completion or near completion of various reptilian genome projects suggests that new studies examining related species with and without TSD could begin to provide additional insight into the evolution of TSD and GSD in vertebrate ancestors.

Morphology and histochemistry of juvenile male American alligator (Alligator mississippiensis) phallus.

Phalli of male crocodilians transfer sperm to female cloaca during sexual intercourse, resulting in internal fertilization. For over a century there have been scientific descriptions of crocodilian phallus morphologies; however, little work has presented detailed cellular-level analyses of these structures. Here we present a histological investigation of the complex functional anatomy of the juvenile male American alligator phallus, including fibrous and vascular erectile structures, a variety of secretory epithelium morphologies, and observed immune cells.

Gonadotropin-induced changes in oviducal mRNA expression levels of sex steroid hormone receptors and activin-related signaling factors in the alligator.

Oviducts respond to hormonal cues from ovaries with tissue proliferation and differentiation in preparation of transporting and fostering gametes. These responses produce oviducal microenvironments conducive to reproductive success. Here, we investigated changes in circulating plasma sex steroid hormones concentrations and ovarian and oviducal mRNA expression to an in vivo gonadotropin (FSH) challenge in sexually immature, five-month-old alligators.

Gene-environment interactions: the potential role of contaminants in somatic growth and the development of the reproductive system of the American alligator.

Developing organisms interpret and integrate environmental signals to produce adaptive phenotypes that are prospectively suited for probable demands in later life. This plasticity can be disrupted when embryos are impacted by exogenous contaminants, such as environmental pollutants, producing potentially deleterious and long-lasting mismatches between phenotype and the future environment. We investigated the ability for in ovo environmental contaminant exposure to alter the growth trajectory and ovarian function of alligators at five months after hatching.

Altered gonadal expression of TGF-β superfamily signaling factors in environmental contaminant-exposed juvenile alligators.

Environmental contaminant exposure can influence gonadal steroid signaling milieus; however, little research has investigated the vulnerability of non-steroidal signaling pathways in the gonads. Here we use American alligators (Alligator mississippiensis) hatched from field-collected eggs to analyze gonadal mRNA transcript levels of the activin-inhibin-follistatin gene expression network and growth differentiation factor 9. The eggs were collected from Lake Woodruff National Wildlife Refuge, a site with minimal anthropogenic influence, and Lake Apopka, a highly contaminated lake adjacent to a former EPA Superfund site.

Phylogenomic analyses reveal the evolutionary origin of the inhibin alpha-subunit, a unique TGFbeta superfamily antagonist.

Transforming growth factor-beta (TGFbeta) homologues form a diverse superfamily that arose early in animal evolution and control cellular function through membrane-spanning, conserved serine-threonine kinases (RII and RI receptors). Activin and inhibin are related dimers within the TGFbeta superfamily that share a common beta-subunit. The evolution of the inhibin alpha-subunit created the only antagonist within the TGFbeta superfamily and the only member known to act as an endocrine hormone.

Altered sex hormone concentrations and gonadal mRNA expression levels of activin signaling factors in hatchling alligators from a contaminated Florida lake.

Activins and estrogens participate in regulating the breakdown of ovarian germ cell nests and follicle assembly in mammals. In 1994, our group reported elevated frequencies of abnormal, multioocytic ovarian follicles in 6 month old, environmental contaminant-exposed female alligators after gonadotropin challenge. Here, we investigated if maternal contribution of endocrine disrupting contaminants to the egg subsequently alters estrogen/inhibin/activin signaling in hatchling female offspring, putatively predisposing an increased frequency of multioocytic follicle formation.

Posthatching development of Alligator mississippiensis ovary and testis.

We investigated ovary and testis development of Alligator mississippiensis during the first 5 months posthatch. To better describe follicle assembly and seminiferous cord development, we used histochemical techniques to detect carbohydrate-rich extracellular matrix components in 1-week, 1-month, 3-month, and 5-month-old gonads. We found profound morphological changes in both ovary and testis.

Influences of sex, incubation temperature, and environmental quality on gonadal estrogen and androgen receptor messenger RNA expression in juvenile American alligators (Alligator mississippiensis).

Gonadal steroid hormone receptors play a vital role in transforming ligand signals into gene expression. We have shown previously that gonads from wild-caught juvenile alligators express greater levels of estrogen receptor 1 (ESR1) than estrogen receptor 2 (ESR2). Furthermore, sexually dimorphic ESR2 mRNA expression (female > male) observed in animals from the reference site (Lake Woodruff, FL, USA) was lost in alligators from the contaminated Lake Apopka (FL, USA).

Morphology and histochemistry of juvenile American alligator (Alligator mississippiensis) nephrons.

Here we present a detailed morphological description of the alligator (Alligator mississippiensis) kidney and nephron. We present a series of histological, histochemical, and immunohistochemical markers that clearly define the seven regions of the alligator nephron. The alligator kidney is composed of many paired (mirrored) lobules on each kidney (lobe).

Increased posthatching mortality and loss of sexually dimorphic gene expression in alligators (Alligator mississippiensis) from a contaminated environment.

A previous study from our laboratory examining development in neonatal alligators from polluted Lake Apopka, Florida, found numerous differences relative to neonates from a reference site, Lake Woodruff National Wildlife Refuge. We postulated that the differences were the result of organizational changes derived from embryonic exposure to environmental contaminants and are related to the poor reproductive success reported in alligators from Lake Apopka. In this study we examine differences in alligators collected as eggs from these two populations and raised under similar conditions for 1 yr.

Alligators, contaminants and steroid hormones.

Steroids are essential for successful reproduction in all vertebrate species. Over the last several decades, extensive research has indicated that exposure to various environmental pollutants can disrupt steroidogenesis and steroid signaling. Although steroidogenesis is regulated by the hypothalamic-pituitary axis, it is also modified by various paracrine and autocrine factors.

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.

Stress and its relation to endocrine function in captive female Siberian sturgeon (Acipenser baeri).

Stress responses to numerous environmental conditions have been studied in a wide range of fish species. Defining the relationship between stress and endocrine function is particularly critical to long-lived species such as sturgeons, whose economic viability relies heavily on proper endocrine function for the production of caviar. In this study, we examined the induced stress response, defined by plasma cortisol and glucose concentrations, and its relationship to plasma 17beta-estradiol, testosterone and 11-ketotestosterone concentrations in cultured female Siberian sturgeon (Acipenser baeri).

Environmental contaminants, fertility, and multioocytic follicles: a lesson from wildlife?

The overall contribution of environmental exposures to infertility is unknown, but a growing scientific database suggests that exposure to various environmental factors, both in utero and neonatally, could dramatically affect adult fertility. Studies of various contaminant-exposed wildlife populations suggest that multiple mechanisms contribute to changes in gonadal development, maturation of germ cells, fertilization, and pregnancy; specifically, the endocrine processes supporting these events. Although great debate and extensive research has occurred during the last decade surrounding fertility, fecundity, and semen quality, much less work has focused on environmental alterations in oocyte development and maturation.