Jon N. Kott

Platelet-derived growth factor B-chain-like immunoreactivity in the developing and adult rat brain

Platelet-derived growth factors (PDGFs) are growth-regulatory molecules that stimulate chemotaxis, proliferation and increased metabolism, primarily of connective tissue cells. In our previous paper, we have demonstrated the ubiquitous localization of PDGF B-chain-containing proteins in neurons and expression of transcripts for PDGF A-chain, B-chain and the two forms of the PDGF receptor in the brains of non-human primates. In the present study, the cellular localization of PDGF B-chain in developing and adult rat brains was analyzed using immunocytochemistry with a PDGF B-chain-specific monoclonal antibody. Intense PDGF B-chain immunoreactivity (PDGFB-I) was distributed around the continuously regenerating primary olfactory neurons at all stages of development from embryo to adult. The major part of PDGFB-I associated with neurons appeared some time after birth and increased with age. PDGFB-I appeared in several nerve fiber systems during earlier stages of development and gradually decreased with age. In conjunction with other data showing the existence of functional PDGF receptor beta-subunits in the neurons, these data suggest a possible role for PDGF B-chain as a neurotrophic or neuroregulatory factor in both developing and mature brains.

Nerve growth factor receptor expression in the young and adult rat olfactory system

Nerve growth factor (NGF) and its receptor (NGFR) are proteins that have a role in the normal development and survival of neurons in the peripheral and central nervous systems. During development, NGF is necessary for outgrowth of axons and establishment of synapses, and NGFR is the transmembrane protein that binds NGF and brings it into the cell. NGF and NGFR expression in the rat olfactory system have been studied previously, and age differences in NGFR are explored further in this study, using immunocytochemistry and immunoelectron microscopy to determine the changes in two different ages: postnatal day 5 and the adult. Dramatic differences were found in the distribution of NGFR immunoreactivity in the olfactory system of each of the two ages studied. Electron microscopy revealed that glial cells were responsible for this immunoreactivity.

Diet selection following area postrema/nucleus of the solitary tract lesions.

Ten adult Long-Evans male rats were offered access to fat, protein and carbohydrate from separate sources. After adaptation to this diet, 5 animals received thermal lesions of the area postrema and adjacent caudal-medial portion of the nucleus of the solitary tract (AP/cmNTS). The remainder were sham-operated. AP/cmNTS lesioned rats ate significantly less and lost more weight than controls during the first postsurgery measurement period (Days 4-13 after lesioning). The decrease of food intake of AP/cmNTS lesioned rats was due to reduced fat consumption. Carbohydrate and protein intakes of lesioned animals did not differ from those of controls. Food intakes and weight changes of lesioned rats did not differ from those of controls during days 14-23 after lesioning. Intake of fat by lesioned animals remained low but was no longer significantly different from that of controls. Carbohydrate and protein intakes of lesioned rats increased slightly but did not differ significantly from those of nonlesioned controls.

Olfactory ensheathing glia and platelet-derived growth factor B-chain reactivity in the transplanted rat olfactory bulb

Using a monoclonal antibody against the B‐chain of platelet‐derived growth factor as a marker, we have examined the behavior of olfactory ensheathing glia in the normal and transplanted rat olfactory bulb. In the normal postnatal olfactory bulb, these glia are found to ensheath the bundles of incoming primary olfactory nerve fibers as well as those in the olfactory nerve layer. Olfactory marker protein antibody was used to identify the olfactory nerve proper. Within the transplant, the same glia: (1) ensheath bundles of both primary olfactory and non‐primary olfactory axons, (2) ensheath axonal bundles deep within the donor tissue, and (3) eventually permit radiation of individual axons from bundles to surrounding neuropil.

Recovery of olfactory behavior. I. Recovery after a complete olfactory bulb lesion correlates with patterns of olfactory nerve penetration.

The olfactory system is an excellent system in which to study issues related to potential functional recovery after a debilitating brain injury. The olfactory system is well-characterized, easily accessible and there are a vast number of studies available from a variety of perspectives. The experimental aim of this research is to examine the anatomical correlates associated with potential behavioral recovery in rats that receive complete olfactory bulb lesions as neonates or as adults. The results show that behavioral recovery occurs only when olfactory nerve penetration of the central nervous system is observed. Further, both olfactory nerve penetration and behavioral recovery are age-dependent phenomena. The olfactory nerve penetration only occurs when the olfactory bulb lesion is performed in neonates. Behavioral recovery of olfactory ability follows a linear trend and reaches near normal levels during the six weeks behavioral testing period. Histological analysis using an antibody for olfactory marker protein (an olfactory nerve-specific marker) reveals two potential candidates for the anatomical pathway responsible for behavioral recovery: olfactory nerve to orbital frontal cortex and olfactory nerve to olfactory peduncle. This report presents evidence that recovery of olfactory ability can occur in the absence of the olfactory bulb if the lesion is performed when the rat is still a neonate.

Area postrema/nucleus of the solitary tract ablations: Analysis of the effects of hypophagia

The effect of hypophagia following lesions of the area postrema and caudal-medial aspect of the nucleus of the solitary tract AP/cmNTS) on body-weight, water intake and preference for palatable diets was examined. Following AP/cmNTS ablation, rats reduced pelleted-food intake to a degree which was sufficient to account for the weight loss and increased water:food ratios observed. Restricting food intakes of intact rats to levels taken by lesioned animals resulted in similar weight losses and increased water:food ratios. When offered both pelleted food and milk, lesioned rats took more calories as milk than did previously food-restricted intact rats. Thus, the hypophagia of AP/cmNTS lesioned rats does not account for their increased preference for milk diets. Lesioned rats ate less high-fat diet than did intact or sham-lesioned controls and did not increase their intakes when this diet was sweetened. At autopsy, retroperitoneal and epididymal fat-pad weights accounted for less of the total body weight of lesioned animals than controls suggesting that body-fat levels are reduced following AP/cmNTS ablation.

Development of olfactory marker protein and tyrosine hydroxylase immunoreactivity in the transplanted rat olfactory bulb

Evidence suggests that tyrosine hydroxylase (TH) expression by juxtaglomerular (JG) neurons of the olfactory bulb (OB) is dependent upon input from primary olfactory neurons (ONs), which are identifiable using immunocytochemical localization (ICC-L) methods for olfactory marker protein (OMP). When the input from the continuously regenerating ONs is temporarily removed (either surgically or chemically), JG cells cease TH production until ON contact is reestablished. We are studying this transneuronal regulation using the rat OB in a transplantation (TX) model. Fetal OBs, labeled in utero with tritiated thymidine, were transplanted (TX) into a site vacated by removal of a neonatal host OB. Host animals were sacrificed at varying periods after TX. Alternate sets of frozen sections were then processed for autoradiography or using ICC-L for TH and OMP. As early as 1 week post-TX, OMP-positive fibers and glomerulus-like structures were seen throughout the TX OB. Despite this extensive and rapid OMP reinnervation, TH expression returned very slowly and the number of TH expressing cells never approached control levels. The reduced TH activity in TXs may be due to failure of JG cells to survive or to develop the correct phenotype under TX conditions. Alternatively, input from ON fibers may only be necessary, but not sufficient, for the expression of TH.

Fetal olfactory bulb transplants send projections to host olfactory cortex in the rat

We are using the rat olfactory system to study developmental details of neurotransplantation. Tritiated [3H]thymidine-labeled fetal olfactory bulbs (OBs), were transplanted immediately into sites from which the neonatal host OB was removed. Subsequently, a small lesion was placed in the region of the transplanted OB and the tissue studied, using degeneration methods and autoradiography. Only OBs with extensive [3H]-label and precise lesions confined to the labeled areas were used. Degeneration was found mainly in the ipsilateral piriform cortex with lesser amounts at other nearby sites. The results demonstrate successfully transplanted donor OBs that send axons to specific and appropriate target areas of the host brain.

Recovery of olfactory behavior. II. Neonatal olfactory bulb transplants enhance the rate of behavioral recovery

Previous experiments in this laboratory have shown that transplants of a fetal olfactory bulb into a neonatal rat are viable and that they establish connections with the olfactory peduncle and olfactory cortex. The focus of this experiment was to investigate the anatomical correlates of any behavioral recovery seen in rats that had one olfactory bulb removed along with an immediate transplant of a fetal olfactory bulb. Anatomical details, such as transplant organization and olfactory nerve repenetration patterns were analyzed using a variety of histological and immunohistochemical techniques. The rats in this experiment showed behavioral recovery of olfactory ability. The recovery rates observed in these animals were compared to two other groups of rats that this laboratory has shown to be behaviorally competent: normal rats and rats with neonatal ablations of the olfactory bulb but no transplant. Although the animals with transplants did not recover to completely normal levels of olfactory ability, they did start behavioral testing in a more behaviorally competent condition than rats with simple neonatal lesions. Anatomical analysis revealed that the transplanted olfactory bulb was heavily penetrated by incoming olfactory nerve fibers but olfactory nerve penetration was not limited to the transplanted olfactory bulb. The extra-bulbar host regions that were penetrated included the orbital frontal cortex and three olfaction-related areas; olfactory cortex, olfactory peduncle and the subependymal cell layer. The olfactory nerve penetration patterns observed beyond the transplant were essentially the same as those observed in rats with only neonatal lesions of the olfactory bulb. Thus, multiple pathways may have contributed to the recovery observed in the rats with olfactory bulb transplants.

Nerve growth factor receptor and the transplanted rat olfactory bulb

Nerve growth factor receptor (NGFR) in the rat olfactory system is developmentally regulated, localizing to the olfactory nerve (ON) in the young rat, and to the olfactory bulb (OB) glomeruli in the adult (Vickland et al. 1991. Brain Res., in press). This pattern of immunoreactivity (IR) may indicate the state of axon growth in the young ON and synaptogenesis in the adult glomeruli. Additional experiments in our laboratory involving lesions to the ON in adult rats have shown a recapitulation of the developmental pattern of expression: NGFR-IR is again found along the ON. Longer survival times after lesioning show the reexpression of the adult pattern of NGFR-IR. This phenomenon was further explored in a transplant (TX) model to determine the changes in NGFR-IR in both the host and TX tissue. A fetal OB labeled with [3H]thymidine is placed into the space created by the removal of the OB of a neonatal rat. After survival times of 1, 2, 8, and 13 weeks, the host animal is sacrificed and the OB TX is processed using monoclonal antibody 192 IgG for NGFR. The host ON shows strong NGFR-IR in TX of 1- and 2-week survival times. In TX survival times of 8 weeks or more, NGFR-IR is observed in glomerulus-like structures. All of these glomerulus-like structures are near groups of neurons in the TX tissue, indicating that they may be functional, with appropriate synapses. Therefore, even though the adult pattern of NGFR-IR takes longer to become established than in normal rats, we have demonstrated that this pattern does so in the TX OB model.