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PubMed2005

Characterization of two avian MHC-like genes reveals an ancient origin of the CD1 family.

Many of the genes that comprise the vertebrate adaptive immune system are conserved across wide evolutionary time scales. Most notably, homologs of the mammalian MHC gene family have been found in virtually all jawed vertebrates, including sharks, bony fishes, reptiles, and birds. The CD1 family of antigen-presenting molecules are related to the MHC class I family but have evolved to bind and present lipid antigens to T cells. Here, we describe two highly divergent nonclassical MHC class I genes found in the chicken (Gallus gallus) that have sequence homology to the mammalian CD1 family of proteins. One of the chicken CD1 genes expresses a full-length transcript, whereas the other has multiple splice variants. Both Southern blot and single nucleotide polymorphism analysis indicates that chicken CD1 is relatively nonpolymorphic. Moreover, cross-hybridizing bands are present in other bird species, suggesting broad conservation in the avian class. Northern analysis of chicken tissue shows a high level of CD1 expression in the bursa and spleen. In addition, molecular modeling predicts that the potential antigen-binding pocket is probably hydrophobic, a universal characteristic of CD1 molecules. Genomic analysis indicates that the CD1 genes are located on chicken chromosome 16 and maps to within 200 kb of the chicken MHC B locus, suggesting that CD1 genes diverged from classical MHC genes while still linked to the major histocompatibility complex locus. The existence of CD1 genes in an avian species suggests that the origin of CD1 extends deep into the evolutionary history of terrestrial vertebrates.

Amino Acid Sequence↗
PubMed1990

Modelling coronary thrombosis from nonanticoagulated human blood in vitro.

The prevalence of ruptured atheromatous plaques underlying the adherent thrombus in the infarct-related coronary arteries, is well documented. In the thrombotic process associated with plaque rupture, hemodynamic forces and the interaction of platelets with exposed collagen fibers play the decisive roles. The shear-induced hemostasis from a nonanticoagulated human blood sample, perfused through polyethylene tubing, was used to simulate rheological changes in the coronary circulation due to plaque disruption. When the hemodynamic conditions of a plaque fissure were mimicked, the sequence of events corresponded to that in vivo: hemostasis (i.e., platelet plug formation in the wall) resulted in the formation of an occlusive thrombus in the lumen of the tubing. Further, the thrombus growth on a collagen fiber, mounted in the lumen of polyethylene tubing through which nonanticoagulated human blood was perfused, was used to mimick the exposure of thrombogenic elements during deep vessel wall injury and the formation of thrombus superimposed on plaque disruption. Morphology of both types of thrombi revealed large numbers of neutrophils and monocytes associated with the platelet mass. The mechanisms of thrombotic reactions were characterized by antagonists and monoclonal antibodies against the platelet activation pathways. Generation of thrombin at an early stage was shown to be the key event and determinant of the final outcome of both thrombotic reactions. It is suggested that the simultaneous measurements of shear-induced hemostasis, clotting, and platelet-collagen interaction from nonanticoagulated human blood provide close experimental approximations to the pathological process of acute coronary syndromes, namely thrombus formation at the site of a disrupted atherosclerotic plaque.

Adolescent↗
PubMed2001

Interaction of processes on different length scales in a bioelastomer capable of performing energy conversion.

This work concerns the aggregation properties of (Gly-Val-Gly-Val-Pro)(251) rec, a polypentapeptide reflecting a highly conserved repetitive unit of the bioelastomer, elastin. On raising the temperature of aqueous solutions above 25 degrees C, this polypeptide was already known to undergo concurrent conformational changes (hydrophobic folding), phase separation, and self-assembly with formation of aggregated three-stranded filaments composed of dynamic polypeptide helices, called beta-spirals. Aggregates obtained from the solution can be shaped into bands that acquire entropic elastic properties upon gamma-irradiation and can perform a variety of energy conversions. Previous studies have shown that aggregation is prompted by the (diverging) critical fluctuations of concentration occurring in the solution, in vicinity of its spinodal line. Here, we present combined circular dicroism (CD) and light scattering experiments, and independent fittings of experimental data to the theoretical spinodal and binodal (coexistence) lines. Results show the following logical and causal sequence of processes: (a) Smooth and progressive conformational changes promoted by concentration fluctuations occurring as temperature is raised "pull down" (in the temperature scale) the instability region of the solution. (b) This further promotes critical fluctuations. (c) The related locally high concentration prompts a further substantial conformational change ending in triple-helix formation and coacervation. (d) This intertwining of processes, covering different length scales (from that of individual peptides to the mesoscopic one of demixed regions), is related to the fact that solvent-induced interactions play a strong role over the entire scale span. These results concur with other recent ones in pointing out that process interactions over many length-scales probably reflect a frequent if not ubiquitous pattern in protein aggregation. This may be highly relevant to the desirable deep understanding of such phenomenon, whose interests cover many fields.

Animals↗
PubMed2002

The complete genome of hyperthermophile Methanopyrus kandleri AV19 and monophyly of archaeal methanogens.

We have determined the complete 1,694,969-nt sequence of the GC-rich genome of Methanopyrus kandleri by using a whole direct genome sequencing approach. This approach is based on unlinking of genomic DNA with the ThermoFidelase version of M. kandleri topoisomerase V and cycle sequencing directed by 2'-modified oligonucleotides (Fimers). Sequencing redundancy (3.3x) was sufficient to assemble the genome with less than one error per 40 kb. Using a combination of sequence database searches and coding potential prediction, 1,692 protein-coding genes and 39 genes for structural RNAs were identified. M. kandleri proteins show an unusually high content of negatively charged amino acids, which might be an adaptation to the high intracellular salinity. Previous phylogenetic analysis of 16S RNA suggested that M. kandleri belonged to a very deep branch, close to the root of the archaeal tree. However, genome comparisons indicate that, in both trees constructed using concatenated alignments of ribosomal proteins and trees based on gene content, M. kandleri consistently groups with other archaeal methanogens. M. kandleri shares the set of genes implicated in methanogenesis and, in part, its operon organization with Methanococcus jannaschii and Methanothermobacter thermoautotrophicum. These findings indicate that archaeal methanogens are monophyletic. A distinctive feature of M. kandleri is the paucity of proteins involved in signaling and regulation of gene expression. Also, M. kandleri appears to have fewer genes acquired via lateral transfer than other archaea. These features might reflect the extreme habitat of this organism.

Base Sequence↗
PubMed1998

Protein engineering of BamHI restriction endonuclease: replacement of Cys54 by Ala enhances catalytic activity.

Chemical modification studies of BamHI endonuclease indicated the importance of the cysteine residue in catalysis [Nath, K. (1981) Arch. Biochem. Biophys, 212, 611-617]. Of the three cysteine residues at positions 34, 54 and 64 in the BamHI endonuclease Cys54 and Cys64 are at the DNA-protein interface. The co-crystal structure of the BamHI-DNA complex, however, does not indicate any role of cysteines either in binding or catalysis. In the context of strong biochemical evidence, Cys54 in BamHI was changed to Ala54 to investigate its role in catalysis. The mutation was carried out by PCR overlap extension, the mutant gene was cloned and characterized by sequencing. The mutant BamHI was expressed and purified to homogeneity and the kinetic parameters (K(M) and kcat) of the wild type and the C54A mutant were determined. The mutation results in up to approximately 40% enhancement of kcat and some increase in K(M). These in vitro results were also supported by in vivo SOS induction assays: the C54A mutant gene under the T7 promoter caused complete lysis in JH139 in absence of T7 RNA polymerase whereas the wild-type gene gave deep blue colonies under the same conditions. The results suggest no direct role of Cys54 in catalysis, but it can influence the catalytic activity through Val57 backbone contact seen in the co-crystal structure.

Alanine↗
PubMed1999

Contact guidance of chick embryo neurons on single scratches in glass and on underlying aligned human skin fibroblasts.

The influence of substratum topography on the morphology and orientation of neurites of chick embryo neurons was studied. Two series of experiments are reported. One concerned the behaviour of growth cones when the axons become contact-guided by the surface texture. The second studied contact guidance of neurites extending on a compact layer of fixed aligned human skin fibroblasts (HSF). It was observed that when the growth cones of sensory neurons isolated from dorsal root ganglions encountered a single scratch in a glass surface (0.1-2 microm in depth and diameter) they turned and continued movement following the axis of the scratch. These neurons became contact-guided as a result of the sequence of events. The growth cone filopodia recognized the irregularity in the substratum surface, whereas the growth cone lamella stabilized contact with the scratch and moved forward along the scratch axis. Scanning electron microscope revealed that the single scratches 150 nm in width and ca. 100 nm deep growth cone filopodia less than 200 nm in diameter could detect and react by turning into them. These filopodia extensions followed the edge of scratches. However, phase contrast and Nomarski's differential interference contrast appeared insufficient for analysis of primary contact guidance of fine growth cone filopodia which themselves are often less than 200 nm. In neuron cultures on fixed aligned HSF, the neuron aggregates assumed spindle-like shapes, and sparsely seeded individual neurons extended axons along the long axes of the fibroblasts. The axons extended significantly further on the fixed underlying fibroblasts than on collagen-covered glass. In crowded cultures of neurons, the cells extended neurites ignoring both the surface anisotropy (the scratches) and the orientation of the aligned fibroblasts. Immunofluorescence staining of neurons with antibodies against neurofilaments made it possible to analyse their shape and orientation on the fibroblasts. Computer-assisted image analysis permitted the observed alignment of the neurites to be characterized quantitatively.

Animals↗
PubMed2002

Normalized cDNA libraries from a porcine model of orthopedic implant-associated infection.

Staphylococcal infections that result from an alteration in a patient's immune response at the surgical site are a major problem in procedures that incorporate biomaterials in trauma surgery and joint replacement. Diagnosis of infection based on pathogen detection is difficult and exacerbated by increasing numbers of partially or totally resistant strains of nosocomial pathogens, particularly Staphylococcus aureus. Expression profiling of a host's cellular immune response could facilitate the identification of the pathways involved in pathogen recognition and eradication and could lead to more rational design of drugs and therapies. To this end, we constructed and characterized ten individually tagged and directionally cloned cDNA libraries from peripheral blood cells (PBC), spleen (Sp), thymus (Th), lymph node (LN), and bone marrow (BM) from immunologically naive and challenged pigs as part of an implant-associated orthopedic model of deep infection. Three of these libraries were normalized at C(0)(t) values 5, 10, 20, and 30. The libraries comprise more than 20 million primary transformants with an average insert length >1.4 kb. Cluster analysis of 7620 ESTs revealed 1029 clusters containing an average of 3.6 sequences and 3846 singletons. Gene discovery is estimated to be approximately 64%. Searches of public databases resulted in 49.3% annotated porcine sequences, of which 22.2% had significant homologies to ESTs from a variety of species, and 28.5% were without a significant match in any public database. We also identified 9.1% ESTs as involved in host cell and organism defense and 11.5% related to cell signaling and communication. These sequences, together with the 28.5% appearing as novel, are of specific interest to the infectious disease process.

Amino Acid Sequence↗
PubMed2006

Crigler-Najjar syndrome type 2.

Crigler-Najjar syndrome is a rare disorder of bilirubin metabolism with two distinct forms: type 1 and type 2. We report three patients with Crigler-Najjar syndrome type 2 (CN-2). All patients had serum bilirubin values higher than 171 micromol/L and deep yellow skin color. The results of other liver function tests, glucose-6-phosphate dehydrogenase activity and hematology tests were normal, and immunologic tests for hepatitis A, B and C were negative, although one patient had slightly elevated alanine aminotransferase level (45 IU/L). Polymerase chain reaction and sequence analysis of the UDP-glucuronosyltransferase 1A1 (UGT1A1) gene revealed a novel homozygous T>A mutation at nucleotide 479 in exon 1 (Val160Glu) of patient 1, a novel homozygous A>G mutation at nucleotide 610 in exon 1 (Met204Val) of patient 2, and a homozygous T>G variation at nucleotide 1456 in exon 5 (Tyr486Asp) plus a heterozygous G>A variation at nucleotide 211 in exon 1 (Gly71Arg/normal) of patient 3. Two of these mutations were novel and variations identified within the coding region of the UGT1A1 gene were considered the cause of CN-2 in all three patients.

Adult↗
PubMedDate not supplied

Postnatal development of substance-P immunoreactivity in the rat superior colliculus.

Immunocytochemical techniques have been used to examine the distribution of substance-P (SP)-labeled neurons in the superior colliculus of rats from birth to adulthood. At birth, there are almost no SP-immunopositive neurons in the tectum. A small number of SP neurons appear over the next several days. However, the vast majority of SP neurons appear between P9 and P10, and by P12 have attained adult-like numbers and distribution. Neurons are confined to the superficial layers of the colliculus, specifically the upper two-thirds of the stratum griseum superficiale (SGS). There is no indication of a differential developmental sequence along rostrocaudal or mediolateral axes. Neuronal types can be distinguished as early as P6 and include horizontal, vertical, and multipolar cells. Substance-P-immunoreactive axons and boutons are also present in the superior colliculus at birth, and are for the most part confined to the deep layers. Boutons are generally of the en-passant type. The density of labeled axons and boutons increases progressively, and by P10 there is an almost adult-like lamination and patchiness. In the adult, labeled axons and boutons are most dense in the stratum opticum and stratum griseum intermedium. Bridges of dorsoventrally oriented labeled axons span the relatively label-poor stratum album intermedium. SP label in the stratum griseum profundum is dense and patchy, and there is also dense label in the stratum album profundum bordering the periaqueductal grey. The role of substance-P-labeled neurons in the superior colliculus is still a matter of speculation. The findings of this study indicate that SP neurons may play a role in intrinsic collicular circuitry.

Animals↗
PubMed2026

Full-length single-cell spatial transcriptomics reveals spatial and cell-type-specific transcript isoforms in the primate brain.

The primate brain exhibits complex RNA alternative splicing heterogeneity crucial for functional complexity, yet systematic spatial isoform characterization has been lacking. We developed Fullscope-seq, a full-length single-molecule large field-of-view spatial transcriptomics sequencing method at single-cell resolution, based on programmed concatenation cDNA for multiple long-read sequencing platforms. Applying Fullscope-seq to the macaque brain, we uncovered thousands of genes exhibiting differential transcript usage (DTU) across cortical layers, cell types and brain regions. Fullscope-seq resolved hundreds of major isoform switches across distinct brain regions and identified DTUs between superficial and deep cortical layers. Cortical layer-specific DTUs showed cell-composition dependence, whereas regional DTUs were regulated according to both cellular composition and spatial contexts. These isoform variations showed substantial enrichment for neuropsychiatric disorder-associated genes and were conserved across platforms and species. Our study establishes a scalable framework for spatial isoform analysis and provides a resource for understanding transcriptomic diversity in complex tissues.

Animals↗
PubMed2000

Crystal structure of human squalene synthase. A key enzyme in cholesterol biosynthesis.

Squalene synthase catalyzes the biosynthesis of squalene, a key cholesterol precursor, through a reductive dimerization of two farnesyl diphosphate (FPP) molecules. The reaction is unique when compared with those of other FPP-utilizing enzymes and proceeds in two distinct steps, both of which involve the formation of carbocationic reaction intermediates. Because FPP is located at the final branch point in the isoprenoid biosynthesis pathway, its conversion to squalene through the action of squalene synthase represents the first committed step in the formation of cholesterol, making it an attractive target for therapeutic intervention. We have determined, for the first time, the crystal structures of recombinant human squalene synthase complexed with several different inhibitors. The structure shows that SQS is folded as a single domain, with a large channel in the middle of one face. The active sites of the two half-reactions catalyzed by the enzyme are located in the central channel, which is lined on both sides by conserved aspartate and arginine residues, which are known from mutagenesis experiments to be involved in FPP binding. One end of this channel is exposed to solvent, whereas the other end leads to a completely enclosed pocket surrounded by conserved hydrophobic residues. These observations, along with mutagenesis data identifying residues that affect substrate binding and activity, suggest that two molecules of FPP bind at one end of the channel, where the active center of the first half-reaction is located, and then the stable reaction intermediate moves into the deep pocket, where it is sequestered from solvent and the second half-reaction occurs. Five alpha helices surrounding the active center are structurally homologous to the active core in the three other isoprenoid biosynthetic enzymes whose crystal structures are known, even though there is no detectable sequence homology.

Alkyl and Aryl Transferases↗
PubMed1992

Oxygen saturation and cardiovascular changes during fibreoptic intubation under general anaesthesia.

Sixty patients who required fibreoptic nasotracheal intubation were studied. Arterial oxygen saturation, arterial blood pressure and heart rate were monitored continuously during fibreoptic intubation under deep halothane anaesthesia. There were significant decreases (p less than 0.001) in arterial blood pressure and heart rate despite administration of intravenous colloid and atropine. Almost one third of the patients (18 out of 60) suffered a decrease in arterial oxygen saturation below 90% during the intubation sequence and in five patients the saturation fell below 80%. The episodes of desaturation were not related to the induction-intubation time or to the grade of laryngeal visibility at direct laryngoscopy.

Adolescent↗
PubMed2003

Fluids from aging ocean crust that support microbial life.

Little is known about the potential for life in the vast, low-temperature (<100 degrees C) reservoir of fluids within mid-ocean ridge flank and ocean basin crust. Recently, an overpressured 300-meter-deep borehole was fitted with an experimental seal (CORK) delivering crustal fluids to the sea floor for discrete and large-volume sampling and characterization. Results demonstrate that the 65 degrees C fluids from 3.5-million-year-old ocean crust support microbial growth. Ribosomal RNA gene sequence data indicate the presence of diverse Bacteria and Archaea, including gene clones of varying degrees of relatedness to known nitrate reducers (with ammonia production), thermophilic sulfate reducers, and thermophilic fermentative heterotrophs, all consistent with fluid chemistry.

Ammonia↗
PubMed2006

Mutations of DNAI1 in primary ciliary dyskinesia: evidence of founder effect in a common mutation.

RATIONALE: Primary ciliary dyskinesia (PCD) is a rare, usually autosomal recessive, genetic disorder characterized by ciliary dysfunction, sino-pulmonary disease, and situs inversus. Disease-causing mutations have been reported in DNAI1 and DNAH5 encoding outer dynein arm (ODA) proteins of cilia. OBJECTIVES: We analyzed DNAI1 to identify disease-causing mutations in PCD and to determine if the previously reported IVS1+2_3insT (219+3insT) mutation represents a "founder" or "hot spot" mutation. METHODS: Patients with PCD from 179 unrelated families were studied. Exclusion mapping showed no linkage to DNAI1 for 13 families; the entire coding region was sequenced in a patient from the remaining 166 families. Reverse transcriptase-polymerase chain reaction (RT-PCR) was performed on nasal epithelial RNA in 14 families. RESULTS: Mutations in DNAI1 including 12 novel mutations were identified in 16 of 179 (9%) families; 14 harbored biallelic mutations. Deep intronic splice mutations were not identified by reverse transcriptase-polymerase chain reaction. The prevalence of mutations in families with defined ODA defect was 13%; no mutations were found in patients without a defined ODA defect. The previously reported IVS1+2_3insT mutation accounted for 57% (17/30) of mutant alleles, and marker analysis indicates a common founder for this mutation. Seven mutations occurred in three exons (13, 16, and 17); taken together with previous reports, these three exons are emerging as mutation clusters harboring 29% (12/42) of mutant alleles. CONCLUSIONS: A total of 10% of patients with PCD are estimated to harbor mutations in DNAI1; most occur as a common founder IVS1+2_3insT or in exons 13, 16, and 17. This information is useful for establishing a clinical molecular genetic test for PCD.

Adolescent↗
PubMed2026

Necrotizing fasciitis associated with Acinetobacter lactucae and Bacillus paramobilis infections in 2 dogs.

Two immunocompetent dogs from separate households were presented to a tertiary referral hospital with soft tissue wounds attributable to severe bacterial infection. A 5-y-old, castrated male Golden Retriever dog (case 1) was presented with extensive hemorrhagic cellulitis of the forelimbs and neck that developed over 48-h and was attributable to Acinetobacter lactucae infection. Additionally, an 8-y-old, spayed female Labrador Retriever-mix dog (case 2) sustained a deep penetrating wound over the left hip that progressed over 24&#x2009;h to necrotizing fasciitis, from which Bacillus paramobilis was isolated. Histopathologic findings in both cases included severe acute, skeletal muscle necrosis and necrosuppurative myofasciitis, dermatitis, and panniculitis with intralesional rod-shaped bacteria. Whole-genome sequencing and phylogenetic analysis of the bacterial isolates revealed numerous cytolytic toxins and other virulence genes carried by the isolates, expanding the profile of these 2 bacteria. To our knowledge, canine cases of A. lactucae and B. paramobilis associated with necrotizing fasciitis have not been reported previously. In both cases, progression from an inciting incident to septic disease was extremely rapid, occurring within 48&#x2009;h in case 1 and 24&#x2009;h in case 2, underscoring the fulminant nature of necrotizing fasciitis.

Animals↗
PubMed2003

Quantitative study of MR T1 and T2 relaxation times and 1HMRS in gray matter of normal adult brain.

OBJECTIVE: To evaluate magnetic resonance (MR) Imaging and (1)H magnetic resonance spectroscopy ((1)HMRS) in the study of normal biochemical process of the brain, as well as differentiation of normal senile brain from cerebral diseases related to senility. METHODS: One hundred and eighty healthy adult volunteers were selected for MR examination and 60 other healthy subjects for (1)HMRS examination. Ages of subjects ranged from 18 to 80 years. They were divided into six age groups. A 0.35 T superconductive MR system was used to perform MR examination. Point resolved spectroscopy sequence was required for (1)HMRS. The metabolites in the spectra included: N-acetylaspartate (NAA), choline compounds (CHO), creatine compounds (CR), myo-inositol (MI), glutamate and glutamine (Glu-n). RESULTS: In 180 cases of MR, the shortest T(2) relaxation time occurred in the deep gray matter within the same age group while the length of T(1) relaxation time was ordered from low to high compared to age groups. T(2) relaxation time decreased as age increased. The peaks, ordered from high to low, were as follows in 60 cases of (1)HMRS: NAA, CR, CHO, MI, Glu-n. The ratios of NAA/CR and Glu-n/CR were higher in the senile age group, while that of MI/CR was lower. The ratio of CHO/CR was increased as age decreased. The ratio of NAA/CR and MI/CR gradually decreased in relation to movement from the anterior to the posterior part of the brain; the ratio of CHO/CR was highest in the occipital cortex. Correlation of T(1) relaxation time and partial metabolite ratios to age were present in gray matter. CONCLUSIONS: Quantitative studies of MR T(1) and T(2) relaxation times and (1)HMRS are essential to evaluation of normal myelinization processes, neuronal integrity and age-related biochemical changes in the brain.

Adolescent↗
PubMed1994

Magnetic resonance imaging in adults presenting with severe acute infectious cellulitis.

BACKGROUND AND DESIGN: Early categorization of some acute soft-tissue infections, such as severe infectious cellulitis (IC) without or with secondary abscess formation, necrotizing fasciitis (NF) or pyomyositis, is frequently difficult. The first one requires only medical treatment, the remaining ones require either surgery or closed drainage. To determine the presence and the extent of these infections early, we have prospectively studied the value of magnetic resonance imaging in patients admitted for IC with local or general criteria of severity. Images were analyzed on a blind basis. Definite diagnosis was obtained by reviewing clinical records and, in most patients, the results of an invasive procedure. RESULTS: Twenty-six patients (56 +/- 23 years old) were included in this study. Among them, 13 received gadolinium-diethylene-triaminepenta-acetic acid intravenously. The final diagnosis was pyomyositis (five patients), NF (three patients), or IC with (seven patients) or without (11 patients) subcutaneous abscess. Images specific for these diseases were best outlined with T2-weighted sequences. In patients with pyomyositis or subcutaneous abscess(es), we observed spindle-shaped or round, well-defined areas of high signal intensity within the muscles or subcutis, respectively. Patients with NF exhibited numerous homogeneous, well-defined dome-shaped areas of hypersignal in the deep hypodermis. In patients with uncomplicated IC, these dome-shaped areas of hypersignal appeared ill-defined, heterogeneous, smaller, thinner, and less numerous than those in patients with NF. CONCLUSIONS: In patients presenting severe IC, magnetic resonance imaging provided an early clue in the diagnosis of pyomyositis, NF, and abscess-complicated IC. By precisely defining the extent of these infections, it helped to plan surgical treatment.

Abscess↗
PubMed1987

The timing of granule cell differentiation and mossy fiber morphogenesis in the opossum.

The timing of developmental events may be important for the orderly formation of neuronal interconnections. In the present study, the timing of granule cell migration is compared with the arrival and maturation of mossy fiber projections. The opossum was chosen as the experimental animal because its protracted postnatal development enables the examination of developmental sequences not as easily recognized in other more commonly used mammalian species. It is shown that all areas that project to the cerebellum as mossy fibers in the adult opossum do so by postnatal day (PD) 30. Their major target, the granule cells begin inward migration from the external germinal layer (EGL) prior to PD 30, but do not form a distinct internal granular layer (IGL) until PD 35. Migrating granule cells penetrate into the IGL deep to granule cells that have begun dendritic differentiation. By PD 50, Golgi impregnations reveal that many granule cells have numerous immature processes, somal spines and dendritic growth cones. After this age these structures are rare and the vast majority of granule cells exhibit short dendrites with digiform endings. Dendritic differentiation subsequent to PD 54 involves an increase in the length of the shaft and the further maturation of terminal digits. Also from Golgi material, immature mossy fiber endings can be identified in the IGL by PD 35 and exhibit mature characteristics at PD 73. Thus, the formation and maturation of granule cell dendrites and their afferents (mossy fibers) occur over an extended period of time (PD 35-73). Moreover, granule cells exhibit a sequence of development similar to that of Purkinje cells: early arrival of their primary afferent projections in the cerebellar anlage; a period of exuberant dendritic growth; and a protracted and overlapping period for dendritic and synaptic maturation.

Animals↗