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Results for “Functional characterization”

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PubMed1994

Structural and functional characterization of the genomic locus encoding the murine beta 2 thyroid hormone receptor.

beta 1 and beta 2 are functional thyroid hormone receptors (TRs) that are generated from the same genomic locus by splicing of a different amino terminus onto a common carboxyl region containing the DNA and hormone binding domains. TR beta 1 is widely expressed whereas TR beta 2 is found primarily in the pituitary gland although low levels of expression have been described in other tissues. To gain insight into the mechanisms governing expression of this complex transcriptional unit, we cloned mouse genomic fragments containing the common carboxyl terminus as well as the unique TR beta 2 amino-terminal sequence that was located at least 25 kilobases upstream. The DNA and ligand binding exons are identical in size and location of their boundaries to those of the human TR beta 1 gene. To determine whether the region 5' of the TR beta 2 amino terminus represented the promoter region, we examined it for sites of transcriptional initiation and for its ability to function as a promoter in TR beta 2-expressing thyrotrope cells. Multiple transcriptional start sites extending over 400 base pairs (bp) were identified with those more proximal showing inhibition by T3. Transcription was not detected more than 400 bp upstream from the putative AUG codon, although initiation downstream of this AUG was demonstrated indicating alternative AUG usage. A fragment containing 500 bp of the TR beta 2 5'-region exhibited preferential promoter activity when transfected into thyrotrope cells that express endogenous TR beta 2. Deletion studies demonstrated that removal of consensus binding sites for the transcription factor Pit-1 resulted in loss of this cell specificity. We therefore conclude that the promoter region responsible for expression of the TR beta 2 isoform in pituitary thyrotropes is distinct from that described for TR beta 1 and is located many kilobases upstream from their common exons.

Amino Acid Sequence↗

PubMed2006

Functional characterization of Drosophila Translin and Trax.

The vertebrate RNA and ssDNA-binding protein Translin has been suggested to function in a variety of cellular processes, including DNA damage response, RNA transport, and translational control. The Translin-associated factor X (Trax) interacts with Translin, and Trax protein stability depends on the presence of Translin. To determine the function of the Drosophila Translin and Trax, we generated a translin null mutant and isolated a trax nonsense mutation. translin and trax single and double mutants are viable, fertile, and phenotypically normal. Meiotic recombination rates and chromosome segregation are also not affected in translin and trax mutants. In addition, we found no evidence for an increased sensitivity for DNA double-strand damage in embryos and developing larvae. Together with the lack of evidence for their involvement in DNA double-strand break checkpoints, this argues against a critical role for Translin and Trax in sensing or repairing such DNA damage. However, Drosophila translin is essential for stabilizing the Translin interaction partner Trax, a function that is surprisingly conserved throughout evolution. Conversely, trax is not essential for Translin stability as trax mutants exhibit normal levels of Translin protein.

Amino Acid Sequence↗

PubMed1999

Expression and functional characterization of the beta-isoform of the folate receptor on CD34(+) cells.

We have investigated the expression and functional competence of folate receptor (FR) isoforms on human hematopoietic cells. Using immunofluorescence and reverse transcriptase-polymerase chain reaction (RT-PCR) methodology, we find that a substantial fraction of low-density mononuclear and CD34(+) cells express both the beta and gamma isoforms of FR. The alpha isoform of FR (the form most commonly found on cancer cells) was surprisingly absent from all hematopoietic cells examined. Compared with KB cells (a human cell line known for its elevated expression of FR-alpha), the abundance of FR-beta on CD34(+) cell surfaces was relatively low (approximately 8% of KB cell levels). Because many antifolates and folic acid-linked chemotherapeutic agents enter malignant cells at least partially via FR endocytosis, it was important to evaluate the ability of FR on CD34(+) cells to bind folic acid (FA). Based on three FR binding assays, freshly isolated CD34(+) cells were found to display no affinity for FA. Thus, regardless of whether steps were taken to remove endogenous folates before receptor binding assays, FR on primitive hematopoietic cells failed to bind 3H-FA, fluorescein isothiocyanate (FITC)-linked FA, or FA-derivatized liposomes. In contrast, analogous studies on KB cells showed high levels of receptor binding for all three FR probes. These studies show that although multipotent hematopoietic progenitor cells express FR, the receptor does not transport significant amounts of FA. Consequently, antifolates and FA-linked chemotherapeutic agents that can be engineered to enter malignant cells exclusively through the FR should not harm progenitor/stem cell function.

Antigens, CD34↗

PubMed1995

Functional characterization of peripheral blood neutrophils in patients with hyperthyroidism.

The thyroid hormones may exert an important effect on neutrophil functions which play a key role in the defence against microbial pathogens. Therefore investigations of the biological functions of neutrophils in peripheral blood of 80 women with hyperthyroidism were performed. The following functions of neutrophils were examined: nitroblue tetrazolium (NBT) reduction ability, phagocytosis, adherence to fibers as well as spontaneous migration of leukocytes and activity of lysozyme in serum. The total number of leukocytes and neutrophils was also determined. The results were compared with those in the control group consisting of 40 healthy women. The data show that in patients with hyperthyroidism, the number of leukocytes and neutrophils was decreased (respectively 4.90 x 10(9) +/- 1.20, 2.70 x 10(9) +/- 0.54 in patients, and 5.40 x 10(9) +/- 1.30, 3.00 x 10(9) +/- 0.74 in the control subjects), index of spontaneous reduction NBT was increased (17.1% +/- 4.5 in patients and 11.1% +/- 4.5 in the control group), index of phagocytosing cells was decreased (63.0% +/- 16.0, 80.3% +/- 14.0 in patients and 72.2% +/- 12.1, 87.4% +/- 12.0 in the control group), an increase in adherence (71% +/- 17 in patients and 58% +/- 10 in the control subjects), and a decrease in the lysozyme activity in undiluted and diluted serum was found (185 IU/ml +/- 85,209 IU/ml +/- 78 in patients and 284 IU/ml +/- 84,352 IU/ml +/- 140 in the control group).

Adult↗

PubMed1996

Structural and functional characterization of vitronectin-derived RGD-containing peptides from human hemofiltrate.

Bioactive peptides derived from the adhesive plasma protein vitronectin are present at submicromolar concentrations in human hemofiltrate of patients with renal diseases and were isolated by a combination of high-efficiency chromatographic steps. The structural and functional properties of these peptides were characterized. Sequencing and mass spectrometry revealed the existence of peptide isoforms (5-6 kDa) which corresponded to the N-terminus (residues 1 to 44-50) of vitronectin. The isolated peptides bound directly to plasminogen-activator inhibitor-1 (PAI-1) and were effective competitors of the interaction of PAI-1 with isolated intact vitronectin or extracellular matrix. These functional properties were indistinguishable from the binding properties of a recombinant fusion protein containing residues 1-52 of vitronectin linked to a portion of glutathione S-transferase, expressed in Escherichia coli. Peptides containing the RGD sequence of vitronectin competed for vitronectin binding to the alpha v beta 3 integrin. No indication for direct growth-factor binding was noted, whereas natural peptides were found associated with PAI-1 as the major binding protein in plasma. These data demonstrate that functionally active vitronectin-derived peptides are released by unknown protease(s) from the mature protein and that these peptides are identical, in terms of activity, to recombinant vitronectin fragments. These natural peptides may interact with active PAI-1 in plasma or at extravascular sites and thereby interfere with established biological functions of intact vitronectin.

Amino Acid Sequence↗

PubMed1999

Molecular cloning and functional characterization of the upstream promoter region of the human p73 gene.

The p73 gene encodes a protein that shares structural and functional homologies with the p53 tumor suppressor protein. To investigate the mechanism of transcriptional regulation of the p73 gene, we isolated a genomic DNA fragment spanning the 5' upstream region of the human p73 gene and characterized the promoter region. Unlike the p53 gene promoter, the human p73 gene promoter contained a putative TATA-box, and did not exhibit any extended homology to the p53 gene. Two CpG islands were located in the 5' upstream region. Transient transfection assays using progressive truncations of the p73 promoter showed that deletion from -119 to +19 relative to exon 1 resulted in a 13- to 20-fold reduction in the p73 promoter activity, suggesting that the elements for basal promoter activity exist in this region, where putative Sp1, AP-2 and Egr-1, 2, 3 sites are located and CpG dinucleotides are especially concentrated.

Base Composition↗

PubMed2005

Structural and functional characterizations reveal the importance of a zinc binding domain in Bloom's syndrome helicase.

Bloom's syndrome (BS) is an autosomal recessive human disorder characterized by genomic instability and a predisposition to a wide variety of cancers. The gene mutated in BS, BLM, encodes a protein containing three domains: an N-terminal domain whose function remains elusive, a helicase domain characterized by seven 'signature' motifs conserved in a wide range of helicases and a C-terminal extension that can be further divided into two sub-domains: RecQ-Ct and HRDC. The RecQ-Ct domain appears essential because two point-mutations altering highly conserved cysteine residues within this domain have been found in BS patients. We report herein that BLM contains a zinc ion. Modelling studies suggest that four conserved cysteine residues within the RecQ-Ct domain coordinate this zinc ion and subsequent mutagenesis studies further confirm this prediction. Biochemical and biophysical studies show that the ATPase, helicase and DNA binding activities of the mutants are severely modified. Structural analysis of both wild-type and mutant proteins reveal that alteration of cysteine residues does not significantly change the overall conformation. The observed defects in ATPase and helicase activities were inferred to result from a compromise of DNA binding. Our results implicate an important role of this zinc binding domain in both DNA binding and protein conformation. They could be pivotal for understanding the molecular basis of BS disease.

Adenosine Triphosphatases↗

PubMed2006

Sympathetic neurotransmission in the rat testicular capsule: functional characterization and identification of mRNA encoding alpha1-adrenoceptor subtypes.

The rat testicular capsule is a thin tissue surrounding the testis, whose precise function is still unknown. We have studied the contractile effects of electrical field stimulation, noradrenaline, and the blockade by antagonists of adrenergic receptors, in order to characterize sympathetic neurotransmission, and adrenoceptor subtypes. In addition, reverse transcription polymerase chain reaction (RT-PCR) assays were made to check for the expression of the three known subtypes of alpha(1)-adrenoceptors. The effects of electrical field stimulation (2 to 20 Hz, 1 ms, 60 V) were almost totally abolished by depletion of neuronal noradrenaline storage with reserpine (10 mg/Kg), but not by the purinergic receptor antagonist suramin (10(-5) M), indicating that noradrenaline, but not ATP, was involved in contractions. The selective alpha(1)-adrenoceptor antagonist prazosin (10(-7) M) was more effective than the selective alpha(2)-adrenoceptor antagonist idazoxan (10(-7) M) to inhibit contractions induced by electrical field stimulation, pointing out a major involvement of alpha(1)-adrenoceptor. When noradrenaline was used instead of electrical field stimulation, it showed a high potency (pD(2)=7.9). Noradrenaline-induced contractions were competitively blocked by the selective alpha(1A)-adrenoceptor antagonists WB 4101 (pA(2)=8.88), phentolamine (pA(2)=8.39) and by the alpha(1B)-adrenoceptor antagonist spiperone (pA(2)=8.57), indicating the presence of functional alpha(1A)- and alpha(1B)-adrenoceptors. In addition, contractions were not blocked by the selective alpha(1D)-adrenoceptor antagonist BMY 7378 (up to 10(-6) M), while selective alpha(2)-adrenoceptor antagonists showed low pA(2) values (yohimbine, 7.25 and idazoxan, 7.49), suggesting a minor role, if any, for alpha(1D)- and alpha(2)-adrenoceptors. To check the proportionate role of alpha(1A)- and alpha(1B)-adrenoceptors, we blocked alpha(1B)-adrenoceptors with chloroethylclonidine (CEC, 30 microM, 45 min), that reduced the maximal effect of noradrenaline by about 60%. The remnant CEC-insensitive noradrenaline contraction was assumed to be unrelated to alpha(1B)-adrenoceptor, and was inhibited by 5-methyl-urapidil (pA(2)=8.94) and by the Ca(2+) channel blocker nifedipine (3 microM), confirming the involvement of alpha(1A)-adrenoceptors. The presence of mRNA encoding alpha(1A)- and alpha(1B)-adrenoceptor was also shown on RT-PCR assays. Unexpectedly, alpha(1D)-transcripts were also detected in these assays. Taken together, our results show that ATP co-transmission could not be detected, and that neurotransmission involves the interaction of noradrenaline with both alpha(1A)- and alpha(1B)-, but not with alpha(1D)- or alpha(2)-adrenoceptor. The fact that the functional alpha(1D)-adrenoceptor could not be detected in spite of the presence of the corresponding mRNA, remains to be investigated.

Acetyl-CoA C-Acyltransferase↗

PubMed2002

Functional characterization of mutations in the GDNF gene of patients with Hirschsprung disease.

Hirschsprung disease (HSCR) is a congenital disorder characterized by the absence of enteric nervous plexuses in hind gut. Ten to forty percent of HSCR patients carry a dominant loss-of-function mutation in the gene encoding the receptor tyrosine kinase RET, a receptor for glial cell line-derived neurotrophic factor (GDNF). Although several mutations have also been found in the GDNF gene of HSCR patients, their impact on GDNF function is unknown. In this study, we have characterized the effect of these mutations on the ability of GDNF to bind and activate its receptors. Although none of the four mutations analyzed appeared to affect the ability of GDNF to activate RET, two of them resulted in a significant reduction in the binding affinity of GDNF for the binding subunit of the receptor complex, GFR(alpha)1. Our results indicate that, although none of the GDNF mutations identified so far in HSCR patients are per se likely to result in HSCR, two of these mutations (i.e. D150N and I211M) may, in conjunction with other genetic lesions, contribute to the pathogenesis of this disease.

Animals↗

PubMed2000

Functional characterization of human sphingosine kinase-1.

Sphingosine kinase catalyzes the phosphorylation of sphingosine to form sphingosine 1-phosphate (SPP), a novel lipid mediator with both intra- and extracellular functions. Based on sequence identity to murine sphingosine kinase (mSPHK1a), we cloned and characterized the first human sphingosine kinase (hSPHK1). The open reading frame of hSPHK1 encodes a 384 amino acid protein with 85% identity and 92% similarity to mSPHK1a at the amino acid level. Similar to mSPHK1a, when HEK293 cells were transfected with hSPHK1, there were marked increases in sphingosine kinase activity resulting in elevated SPP levels. hSPHK1 also specifically phosphorylated D-erythro-sphingosine and to a lesser extent sphinganine, but not other lipids, such as D,L-threo-dihydrosphingosine, N, N-dimethylsphingosine, diacylglycerol, ceramide, or phosphatidylinositol. Northern analysis revealed that hSPHK1 was widely expressed with highest levels in adult liver, kidney, heart and skeletal muscle. Thus, hSPHK1 belongs to a highly conserved unique lipid kinase family that regulates diverse biological functions.

Amino Acid Sequence↗

PubMed2002

Structural and functional characterization of a transcription-enhancing sequence element in the rbcL gene of the Chlamydomonas chloroplast genome.

The structure and function of a transcription-enhancing sequence element in the coding region of the Chlamydomonas reinhardtii rbcL gene was analyzed in Chlamydomonas chloroplast transformants in vivo. The enhancer sequence is contained within a DNA segment extending from position +108 to position +143, relative to the start site of rbcL gene transcription. The sequence remains functional when inverted or when placed 34 bp closer to or 87 bp further downstream of the basic rbcL promoter. However, it does not function from a site about 250 bp downstream of its original location. Besides promoting transcription initiation from the rbcL promoter, the element is able to augment transcription from the promoter of the Chlamydomonas chloroplast atpB gene, but has an inhibitory effect on transcription from the promoter of the chloroplast ribosomal RNA genes. The results suggest that the enhancer-like sequence acts upon transcription initiation in a position-specific and promoter type-specific manner.

Animals↗

PubMed1998

Expression, functional characterization and tissue distribution of a Na+/H+ exchanger cloned from Xenopus laevis oocytes (XL-NHE).

We examined the functional properties of a Na+/H+ exchanger cloned from Xenopus laevis oocytes (XL-NHE) upon stable transfection into PS120 fibroblasts which lack endogenous Na+/H+ exchange. In contrast to untransfected cells, XL-NHE-transfected cells displayed Na+-dependent alkalinization upon acidification with nigericin. XL-NHE activity was inhibited by amiloride, ethylisopropylamiloride, HOE694 [(3-methylsulphonyl-4-piperidinobenzoyl)-guanidine methanesulphonate] and HOE642 [4-isopropyl-3-methylsulphonylbenzoyl)-guanidine methanesulphonate], Ki values being calculated at 5 micromol/l, 25 nmol/l, 300 nmol/l and 180 nmol/l, respectively. The Na+ dependence of pHi recovery was compatible with simple Michaelis-Menten kinetics, the Km for Na+ being 22.0+/-3.2 mmol/l and the Hill coefficient for Na+ being approximately 1. XL-NHE was activated by phorbol ester, whereas forskolin exerted no effect, suggesting the involvement of phospholipase C/protein kinase C signalling pathways rather than protein kinase A signalling pathways in XL-NHE stimulation. Using reverse transcription polymerase chain reaction, XL-NHE message could be detected in various Xenopus tissues including heart, brain, skeletal muscle, reticulocytes, A6-kidney cells and oocytes.

Amiloride↗

PubMed1995

Stable expression and functional characterization of a human cardiac Na+ channel gene in mammalian cells.

In order to develop mammalian cell lines expressing a functional human heart Na+ channel gene (hH1), Chinese hamster ovary (CHO-K1) cells and HeLa cells were transfected with the hH1 gene and the bacterial neomycin (neo) resistance gene. In CHO-K1 cells, direct screening for hH1-positive, G418-resistant colonies by functional patch clamp analysis was complicated due to low-level endogenous expression of a brain-type Na+ channel. Therefore, we developed a stepwise strategy for isolation of cell lines expressing functional hH1 Na+ channels: G418-resistant colonies were sequentially analysed for (1) chromosomal integration of hH1 DNA by PCR, (2) specific hH1 mRNA expression by RT-PCR, (3) hH1 protein production by immunoprecipitation with hH1-specific antisera, and (4) hH1 Na+ channel function by patch-clamp analysis. Using this strategy we obtained two CHO-K1 cell lines which express functional human heart Na+ channels. However, using the same strategy, we were unsuccessful in obtaining functional, hH1-positive HeLa cell lines, even though hH1 mRNA and protein was produced in these cells. The two CHO-K1 cell lines stably express human cardiac Na+ channels which retain normal electrophysiological characteristics with respect to activation and inactivation. In addition, the Na+ channels expressed in these cells are blocked by tetrodotoxin with an IC50 value of 2.5 microM; consistent with known cardiac Na+ channel pharmacology. The density of channels is high enough to permit recording of pseudomacroscopic currents in excised outside-out patches of membrane.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

PubMed1999

Functional characterization of the CFTR R domain using CFTR/MDR1 hybrid and deletion constructs.

To improve our insight into the structure and function of the CFTR R domain, deletion and hybrid constructs in which different parts of the R domain were deleted or replaced by the MDR1 linker domain, and vice versa, were made. Replacement of the linker domain by the R domain did not result in a decrease and replacement of the CFTR R domain by the linker domain did not result in an increase of maturation efficiency, when compared to the respective wild-type proteins. This indicates that the R domain is not responsible for the high degree of degradation observed for CFTR translation products in the ER, but rather the overall structure or sequences located outside the R domain. Replacing the C-terminal part of the R domain (amino acids 780-830) by the MDR1 linker domain resulted in the appearance of PKA-dependent whole cell chloride currents which were not significantly different from wild-type CFTR currents. This might indicate that the PKA sites present in the linker domain are functional and that not the exact sequence of the C-terminal part of the R domain is important, but rather the presence of PKA sites and the length. Moreover, when this hybrid construct was PKC-stimulated, chloride currents were activated. Although these PKC-induced currents were lower than the PKA-induced ones, this again indicates that the linker domain is functional in this hybrid construct. Taken together, these results suggest that the MDR1 linker domain can substitute for part of the regulatory domain of the CFTR protein.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1998

Cobalamin (vitamin B12) biosynthesis: functional characterization of the Bacillus megaterium cbi genes required to convert uroporphyrinogen III into cobyrinic acid a,c-diamide.

The function of individual genes of the Bacillus megaterium cobI operon genes in cobalamin (vitamin B12) biosynthesis was investigated by their ability to complement defined Salmonella typhimurium cob mutants. This strategy confirmed the role of cbiA, -D, -F, -J, -L and cysGA. Furthermore the operon as a whole was used to restore corrin biosynthesis in Escherichia coli, which, although closely related to S. typhimurium, does not possess the CobI pathway. When the B. megaterium cob operon was cloned into a plasmid and transformed into an E. coli strain containing the S. typhimurium cbiP, it conferred upon the host strain the ability to make the cobyric acid de novo. However, cobyric acid synthesis was observed only when the strain was grown anaerobically. Derivatives of the corrin-producing E. coli strain were constructed in which genes of the B. megaterium cob operon had been inactivated. These strains were used to demonstrate that, whereas B. megaterium cbiD, -G and -X are essential for cobyric acid synthesis, the cbiW and -Y genes could be deleted without detriment to cobyric acid production in E. coli.

Bacillus megaterium↗

PubMed2003

Molecular structural and functional characterization of tumor suppressive anti-ErbB-2 monoclonal antibody by phage display system.

To investigate the molecular structural and functional characteristics of tumor-suppressive anti-ErbB-2 monoclonal antibody (mAb) SER4, we performed mAb-gene cloning and epitope mapping by a phage display system. Structural analysis demonstrated that both the heavy chain (HC) and light chain variable regions are highly homologous with the derived germline sequences, while the HC complementarity determining region (HCDR) 3 has a relatively short length and biased amino acid usage. A cloned gene-derived recombinant Fab (rFab) fragment showed antigen binding activity and specificity comparable to the parent mAb. Cross-linking of the rFab fragment with the anti-Fab antibody elicited cell growth inhibition in vitro. These results imply that the cloned genes actually encode the Fab part of SER4. The epitope mimetic peptide (mimotope) isolated by panning a phage-displayed random peptide library against SER4 showed no cross-reactivity with mAbs other than SER4. The mimotope was found to be homologous with (87)AHNQVRQVPLQR(98) in the extracellular domain of ErbB-2 by means of a clustalw search. Since SER4 causes the growth inhibition of ErbB-2 positive cells, the predicted epitope sequence may constitute the putative functional domain of ErbB-2.

Amino Acid Sequence↗

PubMed2005

Functional characterization of T cells in abdominal aortic aneurysms.

Abdominal aortic aneurysms (AAA) exhibit features of a chronic inflammatory disorder. The functional attributes of the T cells in AAA tissue are unclear, with little quantitative or functional data. Using a novel, non-enzymatic method to isolate viable cells from AAA tissue, functional properties of AAA T cells were investigated for the first time. Composition and phenotype of AAA T cells was determined by flow cytometry and verified by immunohistochemistry. Tissue mononuclear cells (MNCs) were cultured in the presence of T-cell mitogens, and cell cycle analysis and cytokine production assessed. Typical cell yield was 4.5 x 10(6) cells per gram of AAA tissue. The majority (58.1+/-5.3%) of haematopoietic (CD45+) cells recovered were CD3+ T cells, B cells comprised 41.1+/-5.7%, natural killer cells 7.3+/-2.5%, and macrophages 2%. Freshly isolated T cells were in resting (G1) state, with 25% expressing the activation-associated cell surface antigens major histocompatibility complex II and CD25. When stimulated in vitro, a significant proportion entered S and G2 phase of the cell cycle, up-regulated CD25, and secreted tumour necrosis factor-alpha, interferon-gamma, interleukin (IL)-5 and IL-6. Despite patient differences, the composition of the AAA inflammatory infiltrate was remarkably consistent, and when re-stimulated ex-vivo T cells produced a stereotypical cytokine response, consistent with the hypothesis that AAA T cells can promote tissue inflammation by secretion of proinflammatory cytokines, and in addition provide signals for B-cell help.

Aged↗

PubMed1994

Modality-dependent modulation of conduction by impulse activity in functionally characterized single cutaneous afferents in the rat.

Cutaneous afferents exhibit changes in excitability after impulse activity that are correlated with functional modality but are independent of axonal diameter, as studied in 39 cold fibers and 51 nociceptors of the rat. Latency of conducted impulses was used to indicate changes in axonal excitability caused by electrical stimulation. Stimuli were applied both at fixed frequencies and at the time intervals of impulses previously recorded during response to natural stimulation. Latency increased following both these forms of electrical stimulation, as well as after natural stimulation of the receptive fields. The latency increase was correlated with the number of impulses and the frequency of the preceding discharge in all of 4 nociceptors and 13 cold fibers studied for this feature. Increase of latency by electrical or natural stimulation led to reduced responsiveness to natural stimulation. The magnitude and time course of latency changes were correlated with fiber modality. In 32 nociceptors the latency increased continuously with time during a stimulus train, whereas in 21 cold fibers there was only an initial increase in latency over the first few seconds, after which the latency remained at a plateau even as the firing response continued. Paralleling this slowing, impulse failure occurred more frequently during repetitive stimulation in both A delta and C nociceptors than in velocity-matched cold fibers of either class. Based on the magnitude of latency increases during stimulus trains at different frequencies, two distinct patterns were discerned in A nociceptors: "Type II" fibers slowed significantly more than "Type I" or cold fibers. The results support the hypotheses (1) that the pattern of latency changes during activity are signatures for the modality in a given fiber; and (2) that endogenous, activity-dependent processes of the axon contribute to adaptation and encoding in cutaneous sensory afferents.

Afferent Pathways↗