蛙科物种背侧褶组织形态特征

背侧褶是无尾两栖类的重要形态特征,但对于其组织结构和功能缺乏了解。本文运用组织学技术,H.E、AB-PAS和Masson三色三种染色方法对蛙科4属的代表物种滇蛙（Dianrana pleuraden）、阔褶蛙（Sylvirana latouchii）、仙琴蛙（Nidirana daunchina）、沼蛙（Boulengerana guentheri）背侧褶和背部皮肤的显微结构进行了系统的比较研究。结果发现：1）蛙科物种的皮肤腺体单元丰富多样,由黏液腺和颗粒腺组成。其黏液腺均为普通黏液腺,按照其分泌物及分泌细胞形态将黏液腺分为Ⅰ型黏液腺和Ⅱ型黏液腺。Ⅰ型黏液腺在沼蛙和仙琴蛙中缺失,Ⅱ型普遍分布。颗粒腺分Ⅰ型颗粒腺、Ⅱ型颗粒腺及特化颗粒腺,Ⅱ型颗粒腺仅见于仙琴蛙,特化颗粒腺仅见于阔褶蛙和沼蛙的背侧褶;2）滇蛙与仙琴蛙背侧褶基本结构与其背部常规皮肤一致,且两种皮肤结构组成也较为相似,在真皮层下都有一层较厚的脂肪层,滇蛙脂肪层厚度约为真皮层厚度的1.52倍,仙琴蛙脂肪层厚度约为真皮层的1.60倍;阔褶蛙与沼蛙背侧褶基本结构也与其常规皮肤一致,且背侧褶结构组成也较为相似,阔褶蛙及沼蛙真皮层内具有单层排列、集中分布的大型特化颗粒腺,其分泌物为两种颗粒物的混合物。3）蛙科中两种背侧褶结构类型代表了两个进化方向,其功能分别为能量储存和反捕,推测其与环境适应进化相关。     

重庆彭水发现务川臭蛙

2022年7月,在重庆市彭水苗族土家族自治县新田镇调查到2只臭蛙,经比较形态特征确认为臭蛙属（Odorrana）物种。基于线粒体16S rRNA基因片段构建的臭蛙属部分物种的贝叶斯和最大似然系统发育树显示,这2只臭蛙和模式产地贵州省务川仡佬族苗族自治县的务川臭蛙（O. wuchuanensis）聚为一支,具有较高的支持率,贝叶斯后验概率为0.99、最大似然超快速引导支持（2 000次重复）自展值为95%。综合形态特征和系统发育分析,确认此2只臭蛙为务川臭蛙,系重庆市首次记录。务川臭蛙分布范围较为狭窄,此前仅记录于贵州省务川县、沿河土家族自治县、荔波县以及湖北省建始县和广西壮族自治区环江毛南族自治县。务川臭蛙在重庆彭水的发现,确认了该物种在武陵山和大娄山脉重庆段的分布。彭水发现的务川臭蛙1雄1雌,雄性体长81.3 mm,大于模式标本的最大体长,大于湖北建始县种群雄蛙体长;雌性体长87.1 mm,在模式标本的体长范围内,小于建始县种群雌蛙体长。线粒体16S rRNA基因的遗传分化分析显示,重庆市彭水苗族土家族自治县的务川臭蛙样本和模式产地的样本没有遗传分化,遗传距离为0。务川臭蛙在重庆市的...     

隆肛蛙皮肤及其腺体的显微结构特征

观察了隆肛蛙(Paa quadranus)皮肤及其腺体的显微结构特点,主要对成体、幼蛙和蝌蚪泄殖腔上方皮肤腺进行了描述和比较。结果表明,隆肛蛙的表皮和真皮内均分布有微血管及黑色素细胞;皮肤腺为泡状腺,腺泡位于真皮浅层的疏松层内,属顶质分泌的粘液腺;雄性成体泄殖腔上方皮肤腺是隆肛蛙的特有结构,属于雄性的第二性征,本文建议称其为肛上腺(supra-anal gland)。文中对肛上腺及其机能、表皮中微血管与皮肤呼吸、黑色素细胞与体温调节等之间的关系进行了讨论。     

大凉疣螈皮肤的组织学观察

对大凉疣螈Tylototriton taliangensis的皮肤进行了显微观察,结果表明,其体表不同部位皮肤的厚度存在一定差异,但基本结构相同.皮肤表面粗糙,表皮角质化程度较高.表皮与真皮相接处毛细血管丰富,毛细血管常向表皮突起,突起处表皮细胞层数减少,背部毛细血管密度大于腹部.皮肤中含有丰富的色素细胞,主要分布在真皮疏松层浅表.真皮中有丰富的皮肤腺:粘液腺体积较小,分布遍及全身,腹侧密度较大;颗粒腺体积较粘液腺大,主要分布在耳区、体背两侧及尾背侧,形成耳后腺、背嵴、尾部颗粒腺区等特殊结构.     

云南昆明地区三种蛙皮肤显微结构的比较

对昆明地区的多疣狭口蛙、昭觉林蛙及黑斑蛙的背腹皮肤切片的显微结构进行了观察和比较,尤其对它们皮肤腺体的类型和功能进行了比较分析.结果 表明:多疣狭口蛙皮肤比昭觉林蛙及黑斑蛙皮肤含有更多的腺体,尤其是颗粒腺比后两者含量更为丰富,并且在皮下存在许多腺体团,背腹皮肤较后者厚.昭觉林蛙皮肤较黑斑蛙的含有更多的颗粒腺和粘液腺.3种蛙的背部皮肤都含有色素细胞,而除了在多疣狭口蛙雌性腹部观察到少量色素细胞外,另两种蛙在腹部皮肤中均未见色素层.相比之下,昭觉林蛙和黑斑蛙皮肤结构较相似,两者同多疣狭口蛙在皮肤腺体分布及数量上差别较大,这也体现了三者不同的生态适应机制.     

务川臭蛙分布区域扩大及其濒危等级再评估

2013年9月在湖北省建始县采集到的2只务川臭蛙 (Odorrana wuchuanensis)标本,为湖北省新纪录。该记录使务川臭蛙的分布区由大娄山脉东段,沿武陵山脉延展至湖北建始县。因此,建议将其濒危等级调低至濒危（Endangered, 缩写为EN）。与模式标本比较,建始县雌蛙标本头体长99.6 mm,远大于模式标本雌蛙的最大记录;成年雄蛙62.8 mm,亦小于模式标本的最小记录;建始雌蛙体侧背皮肤亦有淡色小刺粒,因此,该特征不属于第二性征。     

金佛拟小鲵幼体皮肤的显微结构观察

采用光镜和扫描电镜对金佛拟小鲵(Pseudohynobius jinfo)幼体皮肤进行组织学和形态学观察。金佛拟小鲵幼体皮肤由表皮和真皮构成。不同部位皮肤厚度不同,头部背侧皮肤最薄,其厚度为(45.99±12.77)μm,尾部腹侧的皮肤最厚,其厚度为(95.21±42.72)μm。表皮角质层仅躯干背部和尾部明显,由仍具有一定生理活性的复层扁平上皮细胞构成。皮肤腺体包括黏液腺和颗粒腺。黏液腺广泛分布于身体各个部位的皮肤,颗粒腺呈区域性分布,仅见躯干部和尾部皮肤,其体积大于黏液腺。毛细血管多分布于真皮疏松层腺体周围,与表皮层紧密接触并凸向表皮。色素细胞主要分布于表皮和疏松层的交界处,呈多细胞聚集的状态,形成厚度不一的色素层。     

多疣狭口蛙不同繁殖时期皮肤显微结构观察

采用常规石蜡切片及HE染色,对多疣狭口蛙不同繁殖时期背腹皮肤结构进行显微观察.结果发现:多疣狭口蛙3个不同繁殖时期皮肤结构无明显变化,其背部腺体要比腹部发达.尤其是颗粒腺,比腹部丰富.背部皮肤的色素层也比腹部发达.繁殖前期、繁殖中期粘液腺含量比繁殖后期略微多.     

似鲇高原鳅皮肤与鳍的组织学观察

采用石蜡切片技术及H.E和AB-PAS染色方法,对似鲇高原鳅(Triplophysa siluroides)头部、腹部、背部、侧线部和尾部皮肤结构及胸鳍、腹鳍、背鳍、臀鳍和尾鳍的组织结构进行观察。各部位皮肤均由表皮和真皮构成,真皮包括疏松层和致密层,不同部位皮肤厚度不同。表皮层腹部最厚,为(84.62±10.82)μm,侧线部最薄,为(14.97±3.95)μm,各部位表皮厚度差异显著。表皮层分布着黏液细胞、棒状细胞及味蕾。疏松层头部最厚,为(282.71±70.56)μm,尾部最薄,为(29.07±4.88)μm,该层分布有黑色素细胞、空泡状细胞和颗粒腺,而黏液腺分布于致密层。各部位的鳍均由表皮层、胶原纤维层、胶原下层及鳍条构成,表皮层与皮肤表皮层组成相似,鳍条是矿化的结缔组织。     

版纳鱼螈幼体和成体皮肤的结构

采用光镜和扫描电镜,对我国特有的珍稀濒危两栖动物版纳鱼螈Ichthyophis bannanicus幼体和成体的皮肤进行形态学和组织学观察.版纳鱼螈幼体和成体的皮肤均可分为表皮、真皮疏松层和真皮致密层;皮肤中含有粘液腺和颗粒腺;在不同发育阶段或同一个体的不同部位,其皮肤的各种组成成分在结构和厚度上存在着差异:成体和幼体都是头部的表皮最厚,尾部的最薄;幼体表皮各层细胞分化不明显,几无角化现象,成体表皮的各层细胞分化明显,表层细胞明显角化;成体躯干部皮肤最厚,头部最薄,幼体则是头部皮肤最厚,尾部最薄;幼体和成体的头部皮肤都分布有大量的粘液腺,颗粒腺分布较少;幼体的躯干部皮肤则主要分布着大量颗粒腺,尾部只有颗粒腺,未见粘液腺;成体躯干部和尾部皮肤均分布有大量的颗粒腺和粘液腺.     

The morphology of the dorsal and ventral skin of Triturus karelinii (Caudata: Salamandridae)

In this study, for the first time, the morphology of the dorsal and ventral skin of Triturus karelinii using light microscopy through histochemical methods was described. The skin exhibited basic morphological characteristics of the other urodeles: the epidermis composed of keratinized stratified epithelium with numerous conical protrusions and the dermis subdivided into spongy and compact layers. In the spongious dermis, three distinct types of glands were observed, namely serous, mucous and mixed glands. These glands were alveolar and occurred in both males and females. The morphologies of all three skin glands differed from anurans and other urodeles, having peculiar characteristics. Serous glands exhibited three different appearances. An unusual finding in mucous glands was the different appearance of their granules, showing diverse density after staining with PAS and AB. The histochemical analysis clearly demonstrated the presence of neutral, acid and sulfated mucins in the adenocytes of mucous glands. Mixed glands formed by mucous and serous glands exhibited the same morphological traits of both types of glands. Light microscopic observations revealed that the dorsal and ventral skin showed structural similarities with some minor differences, possibly resulting from their functions.     

The use of retinoids as probes for analyzing morphogenesis of glands from epithelial tissues

Summary Thirty-five years ago Honor Fell and Edward Mellanby were studying effects of high doses of vitamin A on skeletal development in chick embryos when they noticed that a piece of epidermis, accidentally included in an organ culture, had undergone mucous metaplasia. Further studies by Fell and others eventually led to an understanding of the important role of vitamin A in modulating epithelia in vivo. Fifteen years later another organ culture experiment showed me that excess vitamin A could also initiate the morphogenesis of branching and mucus-secreting glands from developing vibrissa follicles in upper lip skin of embryonic mice. Since then our group has shown that induction of this novel structure by naturally occurring retinoids resembles a normal embryonic induction in that it is stage-dependent, time-dependent, and irreversible. Tissue separation and recombination studies showed that isolated upper lip epidermis can form these glands when combined with retinoid-treated upper lip dermis. Untreated mouse epidermis can form similar glands after combination with chick dermis containing higher retinoid levels. The hamster cheek pouch, normally devoid of glandular structures, can also form mucous glands when treated with a retinoid, either in vivo or in vitro. Recombination studies in organ culture have now shown that mesenchyme exposed to retinoid is essential for gland morphogenesis from pouch epithelium. Evidences is accumulating that retinoic acid may even be the active morphogen in some normally developing systems.     

Either chick embryo dermis or retinoid-treated mouse dermis can initiate glandular morphogenesis from mammalian epidermal tissue

Excess retinoids can cause developing mouse vibrissa follicles to be transformed into mucous glands in organ culture. The objective was to test the hypothesis that retinoids act in this system by altering morphogenetic properties of the dermis. After inititation by retinoic acid (RA) in organ culture, glands were shown to develop further in embryonic skin grafted to the chick chorioallantoic membrane (CAM). Recombinants of 12.5 day mouse epidermis with untreated or RA-treated mouse or chick dermis were then grafted to CAM for 7 days. For homospecific recombinants, 13.5 day mouse dermis originated from 11.5 day skin cultured for 2 days, with or without 5.2 microgram/ml RA. For heterospecific recombinants, 12 day dermis came from chick embryos, previously injected with 250 microgram RA. Glands were absent from the homospecific recombinants including untreated mouse dermis, but appeared in 26% of those with RA-treated dermis. Among heterospecific recombinants, 75% of those with RA-treated chick dermis and 29% of those with untreated dermis had glands. Untreated 10-12 day chick skin contained two forms of endogenous vitamin A, retinol (4.5 microgram/g protein) and dehydroretinol (3.7 microgram/g protein), while 13-14 day mouse skin contained only retinol (1.8 microgram/g protein), as shown by high performance liquid chromatography. RA injection increased retinol and dehydroretinol in chick skin, while RA was undetectable. Thus RA can act through mouse dermis to form epithelial glands and through chick dermis to increase the incidence of glands. The glands in recombinants with untreated chick dermis may result from the higher levels of endogenous retinoids in chick skin, compared with mouse skin.     

Regional specificity of anuran larval skin during metamorphosis: dermal specificity in development and histolysis of recombined skin grafts

Summary Skins from back and tail were dissected from tadpoles of Rana japonica prior to resorption of the tail and separated into epidermis and dermis by treatment with neutral protease. Homotypically and heterotypically recombined skins were constructed from the separated epidermis and dermis and transplanted into the tail of the original tadpole. Skin grafts using dermis from tail region degenerated simultaneously with resorption of the tail. However, skin grafts containing dermis from back region survived on the posterior part of the juvenile frog beyond metamorphosis. Furthermore, all epidermis underlaid with dermis from back region formed secretory glands and became flattened epithelia characteristic of adult back skin, regardless of region from which the epidermis came. Even when epidermis isolated from tail skin was cultured inside a back skin graft, the tail epidermis survived forming an epithelial cyst and developed secretory glands. These results suggest that regional specificities of anuran larval skin, i.e., development of back skin and even histolysis of tail skin, are determined by regionally specific dermis. The results also suggest that some of epidermal cells of tail skin are able to differentiate into epithelial cells similar to back skin of the adult under the influence of back dermis.     

Localization of prolactin receptor in the dorsal and ventral skin of the frog (Rana ridibunda)

The dorsal and ventral skin in amphibians plays an important role in osmoregulation. Prolactin hormone is involved in regulation of amphibian skin functions, such as water and electrolyte balance. Therefore, amphibians may be useful as a model for determining the sites of the prolactin receptor. In this study, prolactin receptor was detected in frog dorsal and ventral skin using immunohistochemical staining method. Prolactin receptor immunoreactivity was localized in all epidermal layers except stratum corneum of dorsal skin epidermis, stratum germinativum layer of ventral skin epidermis, myoepithelial cells, secretory epithelium and secretory channel cells of granular glands in both skin regions. The mucous glands and secretory granules of granular glands did not show immunoreactivity for the prolactin receptor. According to our immunohistochemical results, the more widespread detection of prolactin receptor in dorsal skin epidermis indicates that prolactin is more effective in dorsal skin. Presence of prolactin receptors in epidermis points out its possible osmoregulatory effect. Moreover, detection of receptor immunoreactivity in various elements of poison glands in the dermis of both dorsal and ventral skin regions suggests that prolactin has a regulatory effect in gland functions.     

Acceleration of Retinol-Induced Epidermal Mucous Metaplasia by Stimulating the Dermal Adenylate Cyclase-cAMP System in Chick Embryonic Skin: Appearance of cAMP-Dependent Phosphorylated Proteins in Dermis of Retinol-Pretreated Skin after 2 h-Treatment with cAMP

Epidermal mucous metaplasia of cultured 13-day-old chick embryonic tarsometatarsal skin can be induced by culture in medium containing excess retinol (20 μM) for only 8–24 h and then in a chemically defined medium with Bt₂cAMP (0.2–2 mM) and without retinoids or serum for 2 days. In this work, stimulation of the adenylate cyclase-cAMP system in retinol-pretreated skin by forskolin, pertussis toxin, cholera toxin or AIF₄^– was found to accelerate the synthesis of epidermal sulfated glycoprotein (mucin). In skin induced toward mucous metaplasia by retinol, treatment with forskolin for 1 day increased the cAMP content 10-fold in the dermis but only 2-fold in the epidermis over the control levels. The cAMP level of Bt₂cAMP (0.2 mM)-treated skin was 18 times higher in the dermis but rather lower in the epidermis than untreated skin. These results suggest the importance of an adenylate cyclase-cAMP system in the dermis of skin in stimulating mucous metaplasia induced by retinoids. In fact, cAMP-dependent protein phosphorylation was seen only in the dermis of retinol-pretreated skin after 2 h-treatment with cAMP. As no transfer of cAMP from the dermis to the epidermis of forskolin-treated skin was detected, there may be no gap junctional communication between the epidermis and the dermis, while the basement membrane becomes discontinuous during mucous metaplasia.     

Expression pattern of glycoconjugates in the integument of Bufo ictericus (Anuran, Bufonidae): biochemical and histochemical (lectin) profiles

Mucous consists of glycoproteins and proteoglycans produced by specific secretory cells (mucocytes). In anurans the cutaneous mucous is produced by intradermal glands and displays both mechanical and chemical protection functions. Indeed, mucous maintains the integument moist and facilitates gas exchange (cutaneous respiration). In this work, the carbohydrate moiety distribution was investigated in the integument of Bufo ictericus using conventional and lectin histochemistry to describe the pattern of cutaneous glycoconjugate expression, including both secretory and structural proteoglycans. As a preliminary step, the descendent chromatography in Whatmann 1MM paper was undertaken to prepare the histochemical trials involving the lectins. In B. ictericus, the integument exhibits the basic morphological structure found in lower terrestrial vertebrates: the epidermis is a keratinized squamous stratified epithelium supported by spongious and compact layers. The spongy dermis contain secretory portion of both mucous and serous (or poison) glands. The paper chromatography identified galactose, fucose and mannose as characteristic sugar residues. The secretory cells of the mucous gland in the dermis, as well as the interstice between the stratum corneum and the subjacent stratum spinosum in the epidermis exhibit alpha-l-fucose and alpha-galactose residues. The serous glands give no reaction. The alpha-mannose residue was detected in the extracellular matrix of spongious dermis, but not in the dermal glands. The different glycoconjugate location reflects in two glycoconjugates categories: the secretory which participate in the water flow regulation, and the structural which is involved in the dermal maintenance.     

Induction of epidermal mucous metaplasia by culture of recombinants of undifferentiated epidermis and retinol-treated dermis in a chemically defined medium

Epidermal mucous metaplasia of cultured skin is known to be induced by excess retinol. Studies were made on whether retinol affects primarily the epidermis or the dermis during retinol-induced epidermal mucous metaplasia of 13-day-old chick embryonic skin in culture. When recombinants of 13-day-old normal epidermis and retinol-treated dermis were cultured for 7 days in chemically defined medium in the absence of retinol, hormones, and serum, they showed altered epidermal differentiation toward secretory epithelium (mucous metaplasia). Thus retinol acted primarily on dermal cells.     

The nerves in frog skin

In the epidermis of frog skin, most nerves are situated at the top of the basal layer. More superficial nerve fibres are usually adjacent to flask cells; it is concluded that this is not a functional association, but a consequence of the pattern of moulting. There are nerve fibres in the walls of the granular glands; mucous glands appear to have no intrinsic innervation although nerves pass within a short distance of their walls. The smooth muscle bundles of the dermis are innervated, and have a physical attachment to the overlying epidermis.