Detection of pantothenic acid-immunoreactive neurons in the rat lateral septal nucleus by a newly developed antibody

Abstract

Introduction. The available immunohistochemical techniques have documented restricted distribution of vitamins in the mammalian brain. The aim of the study was to develop a highly specific antiserum directed against pantothenic acid to explore the presence of this vitamin in the mammalian brain. Material and methods. According to ELISA tests, the anti-pantothenic acid antiserum used showed a good affinity (10 -8 M) and specificity. The antiserum was raised in rabbits. Using an indirect immunoperoxidase technique, the mapping of pantothenic acid-immunoreactive structures was carried out in the rat brain. Results. Pantothenic acid-immunoreactive perikarya were exclusively found in the intermediate part of the lateral septal nucleus. These cells were generally small, round, fusiform or pyramidal and showed 2-3 long (50-100 µm) immunoreactive dendrites. Any immunoreactive axons containing pantothenic acid were detected. Conclusions. The very restricted anatomical distribution of the pantothenic acid suggests that this vitamin could be involved in some specific neurophysiological mechanisms.

Mapping of folic acid in the children brainstem

“…pantothenic acid, thiamine, riboflavin, pyridoxal) is restricted in the mammalian central nervous system or even a lack of immunoreactivity for some vitamins (e.g. pyridoxine, nicotinamide) has been reported [7][8][9]11]. Here, we did not observe immunoreactivity for RA; however, in rats, the presence of cell bodies containing this vitamin has been demonstrated in two hypothalamic regions (paraventricular hypothalamic nucleus, perifornical region) [10].…”

“…FA, RA, thiamine, riboflavin, vitamin C) has been carried out by using highly specific antisera. These works were performed in rats [10,11], monkeys [3,4,[6][7][8][9] and humans [5]. The presence of immunoreactive fibers containing FA, but not cell bodies, has previously been reported in the adult monkey central nervous system in the following nuclei/regions: geniculate, pulvinar, ventral posteromedial, parafascicular, dorsal mesencephalon and medial thalamus [6].…”

“…In this sense, the distribution of fibers and cell bodies containing folic acid (FA), retinoic acid (RA) (a metabolite of retinol or vitamin A), thiamine (vitamin B 1 ), riboflavin (vitamin B 2 ), nicotinamide (the amide of nicotinic acid or vitamin B 3 ), pantothenic acid (vitamin B 5 ), pyridoxal/pyridoxine (natural forms of vitamin B 6 ) or vitamin C has been reported in the brain of rats, monkeys and humans. Most of the previous studies were performed in the central nervous system of monkeys [3][4][5][6][7][8][9]11]. The knowledge on the anatomical distribution of vitamins leads to gain insight into the role played by these compounds (e.g.…”

Generation of specific antisera directed against D-amino acids: focus on the neuroanatomical distribution of D-glutamate and other D-amino acids

“…D-glutamate is a small molecule with a molecular weight below 1 kDa (MW 147.13). Small molecules are not immunogenic, so they are usually linked through a coupling agent (e.g., aldehydes, carbodiimides, picric acid) to a carrier protein [46,47], since the immune response is triggered by molecules with a mass 1 to 2 kDa, which is the minimum size for stimulating antibody production. Animals receiving the immunogen (D-glutamate-coupling agent-carrier protein) will generate different types of polyclonal antibodies, some of which will recognize the conjugated D-glutamate, while other populations will recognize the carrier protein or the coupling agent.…”

“…1). This is done at 492 nm and the best dilution is obtained when the optical density is around 1 [44,47]. For example, in the case of D-glutamate, several optical densities are obtained for several concentrations of the antibody (dilution 1/40,000: the optical density was 0.5; dilution 1/20,000: 1; dilution 1/10,000: 2; dilution 1/5,000: 3).…”