Our team has been trying to locate and collect the pineal gland in the American bullfrog's brain (Lithobates catesbeianus) for some time now. However, we don't have a standardized protocol for that. Therefore we're not so sure that the tiny tissue we're collecting is actually the pineal gland. Does anyone have any protocols, pictures, or videos for pineal gland collection in amphibians? I really appreciate any help you can provide!
It used to collect the glands of Telmatobius and Hyla frogs (now Hypsiboas). After euthanizing the specimen, I proceeded to extract the lower jaw, exposing the palate. Then, with scissors, I peeled off the epithelium from the mouth, exposing the base of the skull. With scissors, again, I made vertical cuts on the lateral extension of the prootics and anteriorly on the anterior tip of the parasphenoid. After that, I inserted one of the scissors blades through the foramen magnum, horizontally, to make a horizontal cut of the lateral walls of the brain box, on both sides. After that, with forceps, I would lift the ventral covering of the brain cage (parasphenoids and parts of the prootics and exoccipitals). In this way, the ventral base of the brain was exposed. Generally, the hyophysis did not adhere to the bones , and it remained attached to the brain. The pituitary gland is clearly noticeable, differentiating itself from the rest of the nervous tissue, when the material is fresh, due to its great vascularity; it is of oval shape. If it was extracted with fine-tipped forceps, the forceps were passed underneath of the "grain" of the pituitary, which came out easily, tearing the stem of the pars intermedia, but both anterior and posterior lobes remained intact, as he could see later by histological sections. I hope that those memories may help you in any way. If you have questions, please contact me.
You need to confirm it with histology, especially if you publish the results. So do that and you will know :)
It used to collect the glands of Telmatobius and Hyla frogs (now Hypsiboas). After euthanizing the specimen, I proceeded to extract the lower jaw, exposing the palate. Then, with scissors, I peeled off the epithelium from the mouth, exposing the base of the skull. With scissors, again, I made vertical cuts on the lateral extension of the prootics and anteriorly on the anterior tip of the parasphenoid. After that, I inserted one of the scissors blades through the foramen magnum, horizontally, to make a horizontal cut of the lateral walls of the brain box, on both sides. After that, with forceps, I would lift the ventral covering of the brain cage (parasphenoids and parts of the prootics and exoccipitals). In this way, the ventral base of the brain was exposed. Generally, the hyophysis did not adhere to the bones , and it remained attached to the brain. The pituitary gland is clearly noticeable, differentiating itself from the rest of the nervous tissue, when the material is fresh, due to its great vascularity; it is of oval shape. If it was extracted with fine-tipped forceps, the forceps were passed underneath of the "grain" of the pituitary, which came out easily, tearing the stem of the pars intermedia, but both anterior and posterior lobes remained intact, as he could see later by histological sections. I hope that those memories may help you in any way. If you have questions, please contact me.
Another nice tip: being such a small piece of tissue, it requires very little fixing time. When including the pituitary gland in paraffin wax or celloidin, it is a good idea to also include a piece of rose petal, which is very soft for the histological blade and will help to locate the piece at the time of cutting. . Also, the piece can be briefly soaked in eosin prior to embedding to aid visualization.
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