Development Of Acetylcholinesterase Research Articles

Discovery of novel acetylcholinesterase inhibitors through integration of machine learning with genetic algorithm based in silico screening approaches.

Alzheimer's disease (AD) is the most studied progressive eurodegenerative disorder, affecting 40-50 million of the global population. This progressive neurodegenerative disease is marked by gradual and irreversible declines in cognitive functions. The unavailability of therapeutic drug candidates restricting/reversing the progression of this dementia has severed the existing challenge. The development of acetylcholinesterase (AChE) inhibitors retains a great research focus for the discovery of an anti-Alzheimer drug. This study focused on finding AChE inhibitors by applying the machine learning (ML) predictive modeling approach, which is an integral part of the current drug discovery process. In this study, we have extensively utilized ML and other in silico approaches to search for an effective lead molecule against AChE. The output of this study helped us to identify some promising AChE inhibitors. The selected compounds performed well at different levels of analysis and may provide a possible pathway for the future design of potent AChE inhibitors.

SYNTHESIS AND DOCKING STUDIES OF NOVEL TETRAHYDROQUINAZOLINE DERIVATIVE AS PROMISING SCAFFOLD FOR ACETYLCHOLINE ESTERASE INHIBITION

Alzheimer’s disease (AD) is one of the most prevalent neurodegenerative disorder. While pathological hallmarks of this disorder are known, the exact cause of AD remains unclear. Quinazoline was found to be a promising scaffold for the design and development of Acetylcholinesterase (AChE) inhibitors. In this study we report the synthesis of 1'-methyl-3', 4'-dihydro1'H-spiro[cyclopentane-1, 2'-quinazoline] (4) in 73.3% yield. The structure of compound 4 was confirmed with GC-MS, 1H and 13C-NMR. Acetylcholine esterase inhibition was studied virtually with docking into AChE active site and suggests potential use of 4 as a promising scaffold for acetylcholine esterase inhibitor design which might be useful for Alzheimer’s disease.

Peptide-Based Acetylcholinesterase Inhibitor Crosses the Blood-Brain Barrier and Promotes Neuroprotection.

Design and development of acetylcholinesterase (AChE) inhibitor has tremendous implications in the treatment of Alzheimer's disease (AD). Here, we have adopted a computational approach for the design of a peptide based AChE inhibitor from its active site. We identified an octapeptide, which interacts with the catalytic anionic site (CAS) of AChE enzyme and inhibits its activity. Interestingly, this peptide also inhibits amyloid aggregation through its interaction at the 17-21 region of amyloid-beta (Aβ) and stabilizes microtubules by interacting with tubulin as well. Eventually, in the PC12 derived neurons, it shows noncytotoxicity, promotes neurite out-growth, stabilizes intracellular microtubules, and confers significant neuroprotection even upon withdrawal of nerve growth factor (NGF). Further, results reveal that this peptide possesses good serum stability, crosses the blood-brain barrier (BBB), and maintains the healthy architecture of the primary cortical neurons. This work shows discovery of an excellent peptide-based AChE inhibitor with additional potential as a microtubule stabilizer, which will pave the way for the development of potential anti-AD therapeutics in the near future.

Quantifying ligand–receptor interactions for gorge-spanning acetylcholinesterase inhibitors for the treatment of Alzheimer’s disease

There is a need for continued development of acetylcholinesterase (AChE) inhibitors that could prolong the life of acetylcholine in the synaptic cleft and also prevent the aggregation of amyloid peptides associated with Alzheimer’s disease. The lack of a 3D-QSAR model which specifically deconvulates the type of interactions and quantifies them in terms of energies has motivated us to report a CoRIA model vis-à-vis the standard 3D-QSAR methods, CoMFA and CoMSIA. The CoRIA model was found to be statistically superior to the CoMFA and CoMSIA models and it could efficiently extract key residues involved in ligand recognition and binding to AChE. These interactions were quantified to gauge the magnitude of their contribution to the biological activity. In order to validate the CoRIA model, a pharmacophore map was first constructed and then used to virtually screen public databases, from which novel scaffolds were cherry picked that were not present in the training set. The biological activities of these novel molecules were then predicted by the CoRIA, CoMFA, and CoMSIA models. The hits identified were purchased and their biological activities were measured by the Ellman’s method for AChE inhibition. The predicted activities are in unison with the experimentally measured biological activities.

Donepezil hydrochloride: a treatment drug for Alzheimer's disease.

The role of the cholinergic system with respect to cognitive deficits characteristic of Alzheimer's disease (AD) has led to a number of studies focusing on the development of acetylcholinesterase (AChE) inhibitors as a drug for treating this disease. The earliest known AChE inhibitors, namely, physostigmine and tacrine, performed poorly in clinical trials (e.g., poor oral activity, brain penetration, and hepatotoxic liability). Studies were then focused on finding a new type of acetylcholinesterase inhibitor that would overcome the disadvantages of these two compounds. Donepezil hydrochloride inaugurates a new class of AChE inhibitors with longer and more selective action and with manageable adverse effects.

Acetylcholinesterase inhibitors in Alzheimer's disease.

Alzheimer’s Disease (AD) is the most common single cause of dementia in our ageing society. Traditionally thought of as an untreatable degenerative condition, recent advances in drug therapy have challenged this view. The disease is characterised by an insidious decline in cognitive and non-cognitive function. Classically, short and long-term memory is impaired while language skills, concentration and attention are often affected. This results in impaired ability to learn and retain new skills as well as the loss of existing ones. Non-cognitive function is the global term used to describe problems such as depression, agitation, personality changes, delusions and hallucinations. These factors have a significant impact on patient behaviour and a very real impact on the quality of life for both patients and caregivers. Diagnosis of AD is clinically based, and using the NINCDS-ADRDA criteria (Table 1) [1], a diagnosis of probable or possible AD can be made. Definitive diagnosis relies on pathological confirmation, which in the majority of cases is rarely completed. With the development of AD specific treatments, definition of AD from other types of dementia is very important. Table 1 NINCDS-ADRDA Criteria for clinical diagnosis of Alzheimer’s disease. Pathogenesis The pathogenesis of AD has not yet been elucidated. It is widely accepted that a combination of genetic susceptibility factors and environmental triggers are responsible for late onset sporadic AD, the most common form of the disease. An understanding of the disease mechanism remains elusive, and is the key to developing a disease modifying agent. Currently, it is proposed that beta amyloid protein, abnormal tau protein or possibly both play key factors in the development of disease. It has been widely postulated that oxidative damage and a slow inflammatory process are two possible mechanisms involved. As yet, no product with proven disease modifying properties is available, and current treatments offer symptomatic benefit only. The development of acetylcholinesterase (AChe) inhibitor drugs has followed the finding that cholinergic pathways in the cerebral cortex and basal forebrain are compromised in AD [2] and the resultant cholinergic deficit contributes to the cognitive impairment of these patients [3]. Although many believe this ‘cholinergic hypothesis’ to be important, others feel it represents a less significant component of the disease process [4]. Many other neurotransmitters are affected in AD, and the relative importance of each in relation to clinical findings has not been fully elucidated. Initial work focused on the use of acetylcholine precursors, using a similar rationale to dopamine therapy in Parkinson’s disease. A series of small trials using precursors such as choline and phosphatidylcholine showed no reliable improvement in cognitive function, with only 10 out of 43 trials reporting any positive effect [5]. There has been renewed interest in muscarinic agonists drugs, which when first introduced, had major problems with adverse cholinergic effects. Better understanding of the molecular pathology of muscarinic receptors and their subtypes has led to the development of more specific agonists. Drugs such as xanomeline, milameline, and civimeline have reached clinical trials, and the improvements seen in cognitive function are reviewed by Avery et al. [6]. There are also claims that these drugs have disease modifying properties, with effects on APP processing and tau phosphorylation. Muscarinic agonists remain in trial, but have yet to fulfil their potential in AD treatment. The only group of drugs currently licensed for AD treatment is the AChe inhibitors, which act through inhibition of the enzyme acetylcholinesterase (AChe), responsible for the breakdown of ACh in the neural synapse. A meta-analysis of the early AChe inhibitor treatments was encouraging [7] and these proceeded to larger placebo controlled double-blind trials.

Phenotypic characterisation of respecified visual cortex subsequent to prenatal enucleation in the monkey: development of acetylcholinesterase and cytochrome oxidase patterns.

Prenatal bilateral enucleation induces cortex, which normally would have become striate cortex, to follow a default developmental pathway and to take on the cytoarchitectonic appearance of extrastriate cortex (default extrastriate cortex, Dehay et al. [1996] J. Comp. Neurol. 367:70-89). We have investigated if this manipulation influences the cortical expression of acetylcholinesterase (AChE) and cytochrome oxidase (CO). Early enucleation (before embryonic day 81; E81) had only minor effects on the distribution of AChE and CO in the striate cortex. In animals that underwent operation, the striate cortex CO blobs were significantly more closely spaced on the operculum compared with the calcarine. After early enucleation, there was a periodic distribution of CO dense patches in default extrastriate cortex. These CO patches had a center-to-center spacing that was considerably smaller than that of CO stripes in normal area V2, but was somewhat larger than that of the CO blobs in striate cortex. Although the CO stripes characteristic of normal area V2 could not be detected, there were some high-frequency CO patches, similar to those found in default extrastriate cortex. Early enucleation caused a failure to form the transient AChE bands running perpendicular to the striate border, which are normally present in the fetus and early neonate. Late enucleation did not alter AChE expression in extrastriate cortex. The relatively minor effects of early enucleation in the reduced striate cortex contrast with the changes in expression of these enzymes in extrastriate cortex, which accompany large shifts in the location of the striate border. This suggests a massive reorganisation of cortical phenotype in extrastriate cortex.

Postnatal Development of Cortical Acetylcholinesterase-Rich Neurons in the Rat Brain: Permanent and Transient Patterns

The development of acetylcholinesterase (AChE) activity within cortical neurons of the rat brain was investigated using a histochemical method. The fate of these neurons in later stages of development was studied in animals in which AChE within cortical axons (mostly cholinergic) had been depleted by lesions of the cholinergic neurons of the basal forebrain or by injections of diisopropyl fluorophosphate. We designated neurons with medium to high intensity of reaction product as AChEH and neurons with a low intensity of reaction product as AChEL. Four groups of AChEH cortical neurons were detected: (1) AChEH Cajal-Retzius cells were present in layer I at birth (P0) and decreased steadily in number until none could be detected at P17 or thereafter. (2) AChEH neurons within layer VI and underlying white matter were present at P0, peaked in number and staining intensity at P8-P9, showed a moderate decrease in number at P11-P13 and a further decrease into adulthood. (3) AChEH polymorphic intracortical neurons appeared at P3-P4 in deep cortical layers and by P9 were present in layers II-VI. They continued to increase in number through P11-P14 at which time they displayed the adult pattern and were found in all cortical areas. (4) A large population of AChEH pyramidal neurons appeared at P1-P4, peaked at P8-P10 and was no longer visible at P21. In the adult cerebral cortex, few pyramidal neurons displayed AChE activity and these were almost always of the AChEL type. These results indicate that the AChE within cortical neurons is developmentally regulated and that the content of this enzyme helps to differentiate cortical neurons into distinct populations. The transient expression of AChE activity within cortical neurons suggests a role for this enzyme in the development of the cerebral cortex.

Physiological mechanisms mediating enhanced force generation during development and immune sensitization.

We examined the development of acetylcholinesterase (AChase) activity and tracheal smooth muscle (TSM) contraction elicited by acetylcholine (ACh) in a swine model of maturation and a dog model of allergic bronchospasm. Strips of TSM were tethered isometrically at optimal length and responses were expressed as a percentage of the maximum to KCl-substituted perfusate (% KCl). Maximal contraction (ATmax) to ACh in 2-week-old swine (168 +/- 8% KCl) was greater than in 10-week-old swine (142 +/- 2% KCl; p less than 0.02). The AChase inhibitor, physostigmine, augmented ACh-elicited ATmax in 10-week-old (27% increase; p less than 0.01) but not in 2-week-old swine (2% increase; p is NS) and caused a greater increase in sensitivity to muscarinic activation in 2 versus 10 week-old swine (p less than 0.02), thus demonstrating increased contraction of TSM in 2 versus 10-week-old swine, which results at least in part from reduced AChase activity in immature animals. In another study, TSM from ragweed-sensitized dogs demonstrated augmented efficacy to ACh-elicited contraction (180 +/- 6% KCl) compared with TSM from sham-sensitized, littermate controls (163 +/- 4% KCl; p less than 0.05). In the presence of physostigmine, ATmax was not different between ragweed-sensitized and control TSM.(ABSTRACT TRUNCATED AT 250 WORDS)

Maturation of acetylcholinesterase expression in tracheal smooth muscle contraction

We examined postganglionic development of acetylcholinesterase (AChase) activity and tracheal smooth muscle (TSM) contraction elicited by cholinomimetic activation and electrical field depolarization in vitro. Epithelium-intact tracheal strips excised from 21 2-wk-old swine (2ws) and 19 10-wk-old swine (10ws) were tethered isometrically at optimal resting length, and responses were expressed as percent of the maximum to 63 mM potassium-chloride (%KCl). Cumulative concentration-response curves to KCl were equivalent for TSM from 2 and 10ws. However, maximal contraction to ACh in 2ws (168 +/- 8.4 %KCl) was greater than for 10ws (142 +/- 2.3 %KCl; P less than 0.02). Stimulus-response curves (field electrodes; AC source) demonstrated greater sensitivity for TSM in 10ws (stimulus causing 50% of the maximal response = 3.32 +/- 0.13 V in 2ws vs. 2.25 +/- 0.12 V in 10ws; P less than 0.001), indicating that the greater cholinomimetic responsiveness of 2ws did not result from augmented presynaptic nerve conduction. The AChase inhibitor, physostigmine, caused 1) greater sensitivity of responses elicited by electrical field stimulation in 2ws (P less than 0.05) but not in 10ws (P = NS), 2) augmentation of maximal responses to exogenous ACh in 10ws (27% increase; P less than 0.01) but not 2ws (2% increase; P = NS), and 3) a greater increase in sensitivity to cholinomimetic activation in 2ws compared with 10ws (P less than 0.02). These data demonstrate increased cholinergic contraction of TSM in 2 vs. 10ws that results at least in part from reduced AChase activity in the trachea of immature animals.

The development of acetylcholinesterase activity in normal and transplanted superior colliculus in rats.

Histochemical techniques have been used to examine the development of the enzyme acetylcholinesterase (AChE) in normal and transplanted rat superior colliculus (SC). At birth (P0), relatively little AChE activity was found in SC in situ; however, there was a gradual increase in the intensity of AChE staining in the SC over the first 4 postnatal weeks. In the superficial layers, an increase in AChE activity was first seen in rostromedial SC at P6 and was found throughout the upper tectal layers by P10. An increase in AChE activity in the intermediate layers was apparent by P12 and the adult pattern, characterized by periodic bands of AChE staining, was established by P22. In tectal grafts, the development of AChE activity followed a time course similar to that found in normal SC. Mature tectal grafts contained moderate AChE activity with AChE-positive cells scattered throughout the neuropil. There were however, localized, often spherically shaped areas which displayed relatively intense AChE activity. These AChE-dense areas had a characteristic appearance in adjacent sections stained for Nissl or neurofibrils. Significantly, host retinal input, where present, was always restricted to the AChE-dense regions and it seems certain that these areas are homologous to the superficial layers of normal SC. AChE-rich regions were also present, however, in grafts which received no retinal input and in general the pattern of AChE activity in tectal grafts was strikingly similar, irrespective of their location or connections with the host brain. It would appear, therefore, that much of the AChE activity in tectal transplants, and presumably in SC in situ, is intrinsic to that region and not derived from or dependent upon extrinsic innervation.

Presynaptic modulation of activity and molecular forms of acetylcholinesterase in the rat superior cervical ganglion during early postnatal development.

Preganglionic nerve trunk of the rat superior cervical ganglion was transected shortly after birth in order to evaluate the influence of preganglionic nerves on the development of acetylcholinesterase and choline acetyltransferase in ganglionic neurons. In spite of an early decentralization, specific activity of acetylcholinesterase in the ganglion is increasing during the first 3 wk of life until it is about equal to the activity which remains in the superior cervical ganglion decentralized in an adult animal. Thus, the preganglionic nerves, which per se contribute the presynaptic fraction of total ganglionic AChE activity in normal innervated ganglia, apparently exert no significant regulatory effect on the specific activity of the fraction of acetylcholinesterase affiliated with the developing ganglionic cells. However, the absence of innervation during development is strongly reflected in the pattern of acetylcholinesterase molecular forms. The activity of the 16 S molecular form of AChE remains high in the developing superior cervical ganglion, decentralized at birth, in contrast to the substantial absolute and relative decrease of specific activity of this form during development of a normally innervated ganglion. A high proportion of 16 S AChE probably reflects a shift of decentralized immature ganglion nerve cells toward a cholinergic character. In accordance with this assumption, choline acetyltransferase activity in early decentralized ganglia is significantly higher than that in the ganglia decentralized in adult animals.

Development of the multiple molecular forms of acetyl-cholinesterase in chick paravertebral sympathetic ganglia: An in vivo and in vitro study

The development of acetylcholinesterase (AChE) activity and the distribution of this enzyme among its multiple forms was studied in both tissue extracts and dissociated cell cultures of chick paravertebral sympathetic ganglia. In agreement with previous findings, total AChE (expressed either per ganglion or per μg protein) increased in vivo between the time of formation of the paravertebral chain (embryonic day 7; E7) to hatching (E20-E21). After this time, enzyme activity changed much more slowly. Sucrose gradient sedimentation analysis of AChE in ganglia of post-hatching chicks revealed multiple forms of AChE with S values of approximately 6.5, 11 and 19.5 Developmental studies showed that 6.5 S and 11 S forms are present as early as day E7. Much of the pre-hatching increase in total AChE is due to increased levels of the 6.5 S form of the enzyme. By hatching, this form comprised approximately 85–90% of the total AChE activity. In contrast, during the first week after hatching, the activity of the 11 S form increased several-fold while that of the 6.5 S remained approximately unchanged. The 19.5 S form, which is thought to be associated with the synaptic membrane, was not detected prior to day E17 and reached adult levels (2–3% of total AChE activity) by the first week after hatching. Development of AChE was also studied in dissociated cell cultures of embryonic ganglia. Essentially all the AChE activity in such cultures was found to be associated with the neurons. Total AChE activity of cultured E11 ganglia increased in a pattern which was both qualitatively and quantitatively similar to that which occured in vivol. Furthermore, it was found that development of both the 6.5 and 11 S forms of AChE took place in vitro. In cultures of E8, E11, E15, and E19 ganglia, the distribution of activity between the two forms after various times in vitro was similar to that which was found for in vivo ganglia at an equivalent embryonic stage. Such changes were not affected by the elimination of non-neuronal cells from the cultures. Two aspects of in vitro development, however, differed from that which occured in vivo. First, an increase in 11 S AChE did not occur at ages equivalent to the first week post-hatching. Second, the 19.5 S form did not develop (even after several weeks) in cultures of E8, E11 and E15 ganglia, nor was this form (which was removed during dissociation of the ganglia) regenerated in cultures of E19 ganglia. Such findings suggest that the pattern of development of AChE and its multiple forms in chick sympathetic neurons is in part intrinsically programmed into these cells at an early stage of development as well as in part regulated by extrinsic signals that these cells receive from their chemical and cellular environment.

Development of acetylcholinesterase (AChE) staining in human fetal auditory cortex.

In this study acetylcholinesterase (AChE) staining methods have been used to demonstrate the development of the prospective afferent fibres of auditory cortex in human fetuses ranging between 8 and 28 weeks of gestation. Earliest AChE positive staining was found in fetuses at 20--24 weeks in the neuropil of the marginal zone and throughout subplate layer of the auditory cortex. On the basis of this characteristic AChE staining pattern, the auditory cortex may be delineated from surrounding cortical areas. At 24--26 weeks, during intensive lamination of the cortical plate, AchE-positive reaction appears in the deep part of the cortical plate. In the oldest fetuses (28 weeks) a columnar pattern of vertical, darkly stained zones was seen in the middle third of the cortical plate. We conclude that AChE staining is characteristically distributed within cytoarchitectonic compartments and 'synaptic' layers of the developing auditory cortex. The AChE-positive layers coincide with the laminar pattern of synpatogeneis. Thus, AChE-positive reaction during development may serve as a parameter of cortical afferent (thalamic?) innervation of the auditory cortex. In addition, AChE reactivity may indicate the specific transmitter nature of the afferent fibres in the auditory cortex.

Development of acetylcholinesterase during embryogenesis of the ascidian Ciona intestinalis

We have characterized the embryonic muscle cell cholinesterase of the solitary ascidian, Ciona intestinalis (L.). The effects of selective enzyme inhibitors and the inhibition of enzyme activity at high concentrations of substrate suggest that the muscle cell enzyme is an acetylcholinesterase (E.C. 3.1.1.7). After gastrulation and before hatching, acetylcholinesterase activity increased 35- to 40-fold; after hatching (18 hours postfertilization) this activity continued to increase, leveling off at about 36 hours of development. Histochemical observations showed that before hatching acetylcholinesterase was located principally in the muscle cells of the tail and, after hatching, it began to develop in cells of the adult musculature and brain. Inhibition of protein syntnesis by puromycin and of RNA synthesis by actinomycin D, suggest that both protein and RNA synthesis were required for the increase in acetylcholinesterase activity observed in unhatched embryos. Although the continued increase in enzyme activity duirng embryonic development was sensitive to puromycin at all times tested, the actinomycin D sensitivity of this increase was restricted to a discrete time that was completed by about 11 hours of development.

The development of histochemically demonstrable cholinesterases in the rat neostriatum in vivo and in vitro.

The development of acetylcholinesterase (AChE) and non-specific cholinesterase (NsChE) activity was studied in the rat neostriatum by the light and electron microscope using three thiocholine methods. The AChE activity was first demonstrable only in the lateral parts of the nucleus, and during the early postnatal development the most intense activity was in the cell bodies, whilst the typical intense staining of the neuropil of adult animals was seen in two-week-old rats. Two types of AChE-containing cells were observed in the neostriatum of rats younger than two weeks and in cultures of newborn rat neostriatal cells. The neuropil of the cultures showed weak activity in the membranes of thin preterminal processes. In the neuropil of old rats, NsChE activity was present in the membranes of nerve cell processes. The capillary endothelial cells of newborn rats contained both AChE and NsChE. During subsequent development, the AChE activity disappeared, whilst for NsChE no change was seen in the distribution of activity seen in newborn or young adult rats less than three months old.

Effects of acetyl-β-methylcholine on development of acetylcholinesterase and butyrylcholinesterase activities in cultured chick embryonic skeletal muscle

The effect of the presence of methacholine on the development of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activities in cultures of chick embryonic skeletal muscle has been determined. Dissociated thigh muscle cells obtained from 10-day-old chick embryos were placed on collagen-coated cover slips and cultured in Maximow's assembly and transferred to roller tubes. A progressive increase in AChE and BuChE activities occurred during the 10-day culture period. Addition of methacholine to the cuture media increased AChE activity. After irreversible inhibition of AChE and BuChE activities by soman, de novo synthesis of AChE activity was faster in cultures exposed to methacholine than in control cultures. The methacholine-induced development of AChE activity in muscle cultures was blocked by chloramphenicol and puromycin.

Effect of extrinsic neuronal connections on development of acetylcholinesterase and choline acetyltransferase activity in the ventral half of the chick spinal cord

Microsurgical operations were performed on the neural tube of chick embryos of 50–53 hr incubation. The ventral half of the brachial spinal cord subsequently developed in the absence of extrinsic fiber connections. The levels of acetylcholinesterase and choline acetyltransferase activity were measured in the ventral half of the brachial enlargement in both experimental and control embryos from 11–21 days of development. Acetylcholinesterase activity was identical in both experimental and control embryos. The expeirmental values for choline acetyltransferase were less than those of the control: This difference was maximal at 19 days and statistically significant from 15 through 19 days of development. The ontogenetic pattern for each enzyme was similar to that observed for the entire brachial spinal cord. The data suggest that the reduced level of choline acetyltransferase activity in the experimental embryos is a direct result of the operative deficit, i.e., a reduction in the number of cholinergic boutons terminaux in the ventral spinal cord, although possible trophic effects on the motor neuroblast per se cannot be ruled out.

"Trophic" influences of nerve on muscle.

"Trophic" influences of nerve on muscle.

Acetylcholinesterase and cholinesterase activities, protein content and wet weight measures in the rat brain after early hypophysectomy

The development of acetylcholinesterase (AChE) and cholinesterase (ChE) activities, weight measures and protein content were determined for specific brain samples from early hypophysectomized and control female rats at 33–35 days of age. Of the eight brain samples, only two, the somatosensory and ventral cortex, showed significantly less AChE activity in the experimental compared to the control rats. The decrease in the somatosensory AChE activity was postulated to reflect a developmental deficit of sensory-motor function due to the dwarfing of the body mass and surface. The decrease in the ventral cortical sample was interpreted as due to alterations in the development of behavioral and hormonal limbic mechanisms. Early hypophysectomy reduced the level of ChE activity in only one brain region, the hypothalamus. This decrease was thought to be due primarily to alterations in the function of the supraoptic and paraventricular nuclei whose neurosecretory products are normally released from the neurohypophysis. The protein content was not significantly different between the cortical samples from the hypophysectomized and control rats.